System and method for ultra-precise temperature resolution-based digital melting curve analysis and visualization

Abstract

The present invention relates to a digital melting curve analysis and visualization system and method based on ultra-precision temperature resolution that analyzes Tm with improved temperature resolution compared to existing methods based on melting curve data and precisely visualizes it in the form of a histogram. By extracting high-resolution Tm and visualizing histograms through software-based interpolation processing based on data collected from existing equipment, it provides analytical accuracy capable of quantitatively distinguishing minute biological differences such as multiplex targets, single CpG methylation, and microsatellite instability (MSI).

Description

Technology Field

[0001] The present invention relates to a digital melting curve analysis and visualization system and method based on ultra-precision temperature resolution, and more specifically, to a digital melting curve analysis and visualization system and method based on ultra-precision temperature resolution that measures the melting characteristics of nucleic acids in ultra-precision temperature units and analyzes and visualizes them digitally in the fields of molecular biology and gene analysis. Background Technology

[0002] Digital melting curve analysis is a technique used in the fields of molecular biology and genomics to analyze the thermal denaturation characteristics of nucleic acids. By measuring the process in which double-stranded nucleic acids, such as DNA or RNA, separate into single strands as temperature rises, melting curve analysis is utilized for characterizing nucleic acids, detecting mutations, and analyzing gene expression. Conventional digital melting systems typically collect temperature data at intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C. This low-resolution analysis method distorts abrupt shape changes (-dF / dT) on the melting curve through average interval processing, making it difficult to accurately extract Tm and limiting the ability to distinguish subtle differences between samples. In particular, precise molecular changes, such as single CpG methylation or single nucleotide variation, typically exhibit Tm differences of only 0.2–0.3°C; therefore, it is difficult to precisely separate and detect these variations with existing resolutions. In fields where sensitivity and resolution are critical, such as Multiplex PCR, SNP analysis, and Microsatellite Instability (MSI) detection, a temperature resolution of less than 0.1°C is essential. To achieve this, not only software-based interpolation but also hardware-level temperature control, temperature calibration via physical or internal standards (PCR control), and integration with precision analysis applications must be considered in an integrated manner. Furthermore, existing melting analysis is limited to qualitative analysis rather than quantitative information. There is a need for technology that utilizes high-resolution Tm histograms capable of quantitative expression to digitize the temperature-based distribution of individual samples and visualize time-series changes or subtle distribution differences between multiple populations. Consequently, there is an increasing demand for precision analysis technologies capable of simultaneously analyzing biological changes—such as single CpG methylation, MSI, and mutation changes—both quantitatively and qualitatively, and monitoring change trends in the same sample over the long term. The problem to be solved

[0003] The present invention provides a digital melting curve analysis and visualization system and method based on ultra-precision temperature resolution that overcomes the temperature resolution limitations of existing melting curve analysis systems, detects accurate melting points even with minute temperature changes through ultra-precision temperature measurement, and enhances analysis efficiency by intuitively visualizing complex melting curve data. means of solving the problem

[0004] According to one embodiment, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include at least one of a data input module, a temperature resolution correction module, and a high-resolution Tm data processing module. The data input module may use raw melting data collected from a real-time digital PCR platform as input values. The temperature resolution correction module may convert the existing measured temperature intervals into a high-resolution curve. The high-resolution Tm data processing module may precisely extract the interpolated Tm data.

[0005] According to another embodiment, the data input module may include a real-time digital PCR platform interface module that collects data in conjunction with a real-time digital PCR platform. This interface module can directly receive and process data from the digital PCR platform and utilize it as input data for ultra-precision temperature resolution-based digital melting curve analysis.

[0006] According to another embodiment, the temperature resolution correction module may include a linear interpolation processing module that converts existing measurement data at intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C into a high-resolution curve of 0.1°C or less by generating additional temperature points using a linear interpolation method. This linear interpolation processing module can provide finer temperature resolution by calculating values ​​between measured data points.

[0007] According to another embodiment, the temperature resolution correction module may include a derivative curve generation module that improves the precision of Tm extraction by smoothly generating the first derivative (-dF / dT) curve of each well. This derivative curve generation module can provide more precise analysis results by accurately calculating the rate of change of the melting curve.

[0008] According to another embodiment, the high-resolution Tm data processing module may include a Tm data segmentation processing module that generates a histogram by segmenting the x-axis in units of 0.1℃. Through this segmentation processing, temperature data can be analyzed and visualized more finely.

[0009] According to another embodiment, the high-resolution Tm data processing module may include a well count calculation module that displays the number of wells having the corresponding Tm on the y-axis. This well count calculation module enables visual analysis of the distribution of the digital melting curve by calculating the number of wells having the corresponding Tm value at each temperature point and displaying them on the y-axis.

[0010] According to another embodiment, the high-resolution Tm data processing module may include a well count calculation module that represents the number of wells with a corresponding Tm on the y-axis. By calculating the frequency of wells belonging to a specific Tm range, this well count calculation module can perform quantitative analysis on the concentration of a target.

[0011] According to another embodiment, the high-resolution Tm data processing module may include a high-resolution visualization module that visualizes Tm distributions that are close to each other with a resolution 3 to 5 times higher than the existing resolution, making it possible to clearly distinguish between them. This high-resolution visualization module can express the analysis results of the digital melting curve more accurately based on ultra-precision temperature resolution.

[0012] According to another embodiment, the high-resolution Tm data processing module may include a PCR control peak reference alignment module that sets the PCR control peak used in the melting analysis as a reference value and reference-calibrates the Tm values ​​of the entire sample based on this reference. This PCR control peak reference alignment module enables more accurate comparison of Tm values ​​between various samples.

[0013] According to another embodiment, the PCR control peak reference alignment module can consistently adjust the relative position relative to the reference point when comparing data before and after alignment. Through such consistent adjustment, the alignment accuracy of the data can be improved and the reliability of the analysis results can be increased.

[0014] According to another embodiment, the system may further include an analysis software module, which is a software module that executes a high-resolution correction algorithm and a visualization UI. This analysis software module extends the capabilities of the ultra-precision temperature resolution-based digital melting curve analysis and visualization system to enable high-resolution correction and visual representation of data.

[0015] According to another embodiment, the analysis software module may include a data smoothing module that processes data smoothly to improve analysis accuracy. This data smoothing module serves to remove noise from data acquired in an ultra-precision temperature resolution-based digital melting curve analysis and visualization system and to improve the quality of the data.

[0016] According to another embodiment, the analysis software module may include a noise removal module that improves data quality by removing noise from the data. This noise removal module plays a role in enhancing the accuracy and reliability of data in a digital melting curve analysis and visualization system based on ultra-precision temperature resolution.

[0017] According to another embodiment, the analysis software module may include a threshold filter module that filters data based on a set threshold. This threshold filter module can remove unnecessary data during the analysis process or select and process only data that satisfies specific conditions. Through threshold filtering, the quality of the data can be improved and the accuracy of the analysis results can be increased.

[0018] According to one embodiment, a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include at least one of a data input step, a temperature resolution correction step, and a high-resolution Tm data processing step. The data input step may receive raw melting data collected from a real-time digital PCR platform. The temperature resolution correction step may convert the measured temperature data into high-resolution temperature data. The high-resolution Tm data processing step may precisely extract the interpolated Tm data.

[0019] According to another embodiment, the data input step may include a real-time digital PCR platform data interface step that collects data in conjunction with a real-time digital PCR platform. This allows for the efficient collection of data for digital melting curve analysis and visualization based on ultra-precision temperature resolution.

[0020] According to another embodiment, the temperature resolution correction step may include a linear interpolation application step that generates additional temperature points using a linear interpolation method based on the measured temperature intervals and converts them into a high-resolution curve of 0.1°C or less. Through this linear interpolation application, ultra-precision temperature resolution can be achieved to improve the analysis accuracy of the digital melting curve.

[0021] According to another embodiment, the temperature resolution correction step may include a derivative curve generation step that improves the precision of Tm extraction by smoothly generating the first derivative (-dF / dT) curve of each well. Through such derivative curve generation, the analysis accuracy of the digital melting curve can be increased.

[0022] According to another embodiment, the high-resolution Tm data processing step may include a Tm data segmentation step that generates a Tm data histogram by segmenting the x-axis into temperature units smaller than the temperature units of the raw input data. Through this segmentation process, the precision can be increased by analyzing the data in finer temperature units.

[0023] According to another embodiment, the high-resolution Tm data processing step may include a well count calculation step that represents the number of wells with the corresponding Tm on the y-axis of a histogram. Through this well count calculation, the distribution of wells where melting occurred at a specific temperature can be accurately identified.

[0024] According to another embodiment, the high-resolution Tm data processing step may include a high-resolution visualization step that has a resolution 3 to 5 times higher than the existing resolution and visualizes Tm distributions that are close to each other so as to be clearly distinguishable. Through this high-resolution visualization, close Tm distributions that were difficult to distinguish with existing methods can be analyzed by distinguishing them more clearly.

[0025] According to another embodiment, the high-resolution Tm data processing step may include a PCR control peak reference alignment step, which sets the PCR control peak used in the melting analysis as a reference value and reference-calibrates the Tm values ​​of the entire sample based on this reference value to reduce errors between experiments or equipment and ensure quantitative accuracy for comparison between data. Through this PCR control peak reference alignment step, variations that may occur in various experimental environments can be minimized, and more accurate analysis results can be obtained.

[0026] According to another embodiment, the method may further include an analysis software execution step for executing a high-resolution correction algorithm and a visualization UI in the analysis software. Such analysis software execution can improve the efficiency of the ultra-precision temperature resolution-based digital melting curve analysis and visualization method.

[0027] According to another embodiment, the analysis software execution step may include a data smoothing step that processes data smoothly to improve analysis accuracy. This data smoothing step can contribute to increasing the accuracy of the ultra-precision temperature resolution-based digital melting curve analysis and visualization method.

[0028] According to another embodiment, the analysis software execution step may include a noise removal step that improves data quality by removing noise generated in the data. Through this noise removal step, the accuracy of the digital melting curve analysis based on ultra-precision temperature resolution can be increased.

[0029] According to another embodiment, the analysis software execution step may include a threshold filtering step that filters data based on a set threshold. Through this threshold filtering, unnecessary data can be removed and only meaningful data can be selectively processed during the process of digital melting curve analysis and visualization based on ultra-precision temperature resolution.

[0030] According to another embodiment, by software-based interpolation processing based on data collected from existing equipment to extract high-resolution Tm and visualize histograms, analytical accuracy capable of quantitatively distinguishing minute biological differences such as multiplex targets, single CpG methylation, and Microsatellite Instability (MSI) can be provided. Effects of the invention

[0031] The present invention detects accurate melting points even with minute temperature changes through ultra-precision temperature measurement and provides analysis efficiency by intuitively visualizing complex melting curve data.

[0032] The present invention provides analytical accuracy capable of quantitatively distinguishing minute biological differences, such as multiplex targets, single CpG methylation, and Microsatellite Instability (MSI), by software-based interpolation processing based on data collected from existing equipment to extract high-resolution Tm and visualize histograms. Brief explanation of the drawing

[0033] FIG. 1 is a conceptual block diagram of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. FIG. 2 is a conceptual block diagram of a data input module according to an example of an embodiment of the present invention. FIG. 3 is a conceptual block diagram of a temperature resolution correction module according to an example of an embodiment of the present invention. FIG. 4 is a conceptual block diagram of a temperature resolution correction module according to an example of an embodiment of the present invention. FIG. 5 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. FIG. 6 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. FIG. 7 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. FIG. 8 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. FIG. 9 is a conceptual block diagram of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. FIG. 10 is a conceptual block diagram of an analysis software module according to an example of an embodiment of the present invention. FIG. 11 is a conceptual block diagram of an analysis software module according to an example of an embodiment of the present invention. FIG. 12 is a conceptual block diagram of an analysis software module according to an example of an embodiment of the present invention. FIG. 13 is a drawing illustrating an example of a melting curve correction method according to an embodiment of the present invention. FIG. 14 is a diagram illustrating an example of a first derivative (-dF / dT) based peak curve according to one embodiment of the present invention. FIG. 15 is a diagram illustrating the effect of a histogram-based high-resolution Tm analysis and normalization technique according to an embodiment of the present invention compared with a conventional method. In FIG. 15A, the upper graph is a Tm histogram generated based on conventional resolution (approx. 0.5°C units), and the lower graph is the result of analyzing the same data by applying a high-resolution normalization algorithm of 0.1°C units according to the present invention. FIG. 15B is a diagram presenting an example in which the Tm values ​​of the entire sample are normalized based on the representative value of the PCR control (Tm standard) in addition to the high-resolution normalization analysis results of FIG. 15A. In FIG. 16, FIG. 16A is a diagram illustrating a comparative example of RT-qPCR-based multiplex melting curve analysis as background technology of the present invention. FIG. 16B is a diagram showing the results of digital PCR-based melting curve analysis according to the present invention. FIG. 17 is a conceptual diagram illustrating the design of a melting curve analysis for a specific gene promoter according to an embodiment of the present invention. FIG. 18 is a diagram illustrating primer positions and methylation target regions designed based on the CpG island sequence within the promoter of the CDH13 gene as a specific example of the analysis design shown in FIG. 17 according to one embodiment of the present invention. FIG. 19 is a diagram quantitatively showing the results of a digital melting analysis performed based on the design of FIG. 17 and FIG. 18 according to an embodiment of the present invention. FIG. 20 is a conceptual diagram illustrating the progression of methylation histogram analysis and abnormal DNA methylation according to an embodiment of the present invention. FIG. 20A shows the concept of a histogram generated by digital melting analysis of CpG regions of a healthy individual, and FIG. 20B ( FIG. 20b, FIG. 20c) visualizes the pathological progression of abnormal DNA methylation induced by external physical, chemical, and biological stimuli step by step. FIG. 21 is a conceptual diagram illustrating a method for detecting microsatellite instability (MSI) based on digital melting curve analysis and an example thereof according to an embodiment of the present invention. FIG. 21A is a diagram explaining the method for detecting MSI based on digital melting curve analysis according to an embodiment of the present invention, and FIG. 21B is a diagram showing the pathological concept of MSI and an example of detection through digital melting analysis based on the present invention. FIG. 22 is a conceptual flowchart of a digital melting curve analysis and visualization method based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. FIG. 23 is a conceptual flowchart of a data input step according to an example of an embodiment of the present invention. FIG. 24 is a conceptual flowchart of a temperature resolution correction step according to an example of an embodiment of the present invention. FIG. 25 is a conceptual flowchart of a temperature resolution correction step according to an example of an embodiment of the present invention. FIG. 26 is a conceptual flowchart of a high-resolution Tm data processing step according to an example of an embodiment of the present invention. FIG. 27 is a conceptual flowchart of a high-resolution Tm data processing step according to an example of an embodiment of the present invention. FIG. 28 is a conceptual flowchart of a high-resolution Tm data processing step according to an example of an embodiment of the present invention. FIG. 29 is a conceptual flowchart of a digital melting curve analysis and visualization method based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. FIG. 30 is a conceptual flowchart of the analysis software execution step according to an example of an embodiment of the present invention. FIG. 31 is a conceptual flowchart of the analysis software execution step according to an example of an embodiment of the present invention. FIG. 32 is a conceptual flowchart of the analysis software execution step according to an example of an embodiment of the present invention. Specific details for implementing the invention

[0034] Definition of Terms

[0035] In this specification, "melting temperature (Tm)" refers to the point in the transition region where double-stranded nucleic acid denatures into single strands as the temperature rises, at which the rate of change of the fluorescence signal (based on the first derivative) is maximum. This is generally determined by the nucleotide sequence, length, GC content, etc. of the nucleic acid, and is utilized for nucleic acid characterization, mutation detection, SNP classification, etc. Hereinafter, it will be abbreviated as "Tm".

[0036] In this specification, the term “melting curve” (also referred to as “melting curve”) refers to a curve generated by measuring changes in fluorescence signals during the process in which a nucleic acid double strand dissociates into a single strand while the temperature is gradually increased. Hereinafter, it will be abbreviated as “melting curve”.

[0037] In this specification, the term “PCR control peak” refers to the Tm of the reference substance included in the experiment and serves as a reference point for correcting the Tm value of the entire sample. The Tm position of the PCR control peak serves as the standard for reference calibration. Hereinafter, it will be abbreviated as “PCR control peak”.

[0038] In this specification, “binning” refers to a process of dividing continuous Tm data into specific temperature intervals (e.g., 0.1°C) and calculating the frequency for each interval, and is a preprocessing technique for generating histograms. Hereinafter, it will be abbreviated as “binning” in this specification.

[0039] In this specification, the term "well" refers to a microreaction compartment arranged regularly on a microfluidic chip or microplate, and is a unit space capable of individually performing nucleic acid amplification (PCR) or melting reactions. Generally, it is designed to independently hold small sample volumes of less than a microliter, and each well can maintain an independent thermal and chemical environment during the experiment. Hereinafter, it will be abbreviated as "well."

[0040] In this specification, "raw melting data" refers to a set of time or temperature-ordered measurements of fluorescence signals according to temperature change, directly obtained from a digital melting analysis device to form a melting curve. This data is subsequently processed through first derivative analysis (-dF / dT), interpolation, normalization, etc., and is used for analysis such as final Tm extraction and histogram visualization. Hereinafter, this specification will be abbreviated as "raw melting data."

[0042] The present invention relates to a digital melting curve analysis and visualization system and method based on ultra-precision temperature resolution, and more specifically, to a digital melting curve analysis and visualization system and method based on ultra-precision temperature resolution that measures the melting characteristics of nucleic acids in ultra-precision temperature units and analyzes and visualizes them digitally in the fields of molecular biology and gene analysis.

[0043] According to one embodiment, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include at least one of a data input module, a temperature resolution correction module, and a high-resolution Tm data processing module. The data input module may use raw melting data collected from a real-time digital PCR platform as input values. The temperature resolution correction module may convert the existing measured temperature intervals into a high-resolution curve. The high-resolution Tm data processing module may precisely extract the interpolated Tm data.

[0044] According to another embodiment, the data input module may include a real-time digital PCR platform interface module that collects data in conjunction with a real-time digital PCR platform. This interface module can directly receive and process data from the digital PCR platform and utilize it as input data for ultra-precision temperature resolution-based digital melting curve analysis.

[0045] According to another embodiment, the temperature resolution correction module may include a linear interpolation processing module that converts existing measurement data of a temperature interval of 0.2°C or more into a high-resolution curve of 0.1°C or less by generating additional temperature points using a linear interpolation method. This linear interpolation processing module can provide finer temperature resolution by calculating values ​​between measured data points.

[0046] According to another embodiment, the temperature resolution correction module may include a derivative curve generation module that improves the precision of Tm extraction by smoothly generating the first derivative (-dF / dT) curve of each well. This derivative curve generation module can provide more precise analysis results by accurately calculating the rate of change of the melting curve.

[0047] According to another embodiment, the high-resolution Tm data processing module may include a Tm data segmentation processing module that generates a histogram by segmenting the x-axis into units of 0.1℃ or less. Through this segmentation processing, temperature data can be analyzed and visualized more finely.

[0048] According to another embodiment, the high-resolution Tm data processing module may include a well count calculation module that displays the number of wells having the corresponding Tm on the y-axis. This well count calculation module enables visual analysis of the distribution of the digital melting curve by calculating the number of wells having the corresponding Tm value at each temperature point and displaying them on the y-axis.

[0049] According to another embodiment, the high-resolution Tm data processing module may include a high-resolution visualization module that visualizes Tm distributions that are close to each other with a resolution 3 to 5 times higher than the existing resolution, making it possible to clearly distinguish between them. This high-resolution visualization module can express the analysis results of the digital melting curve more accurately based on ultra-precision temperature resolution.

[0050] According to another embodiment, the high-resolution Tm data processing module may include a PCR control peak reference alignment module that sets the PCR control peak used in the melting analysis as a reference value and reference-calibrates the Tm values ​​of the entire sample based on this reference. This PCR control peak reference alignment module enables more accurate comparison of Tm values ​​between various samples.

[0051] According to another embodiment, the PCR control peak reference alignment module can consistently adjust the relative position relative to the reference point when comparing data before and after alignment. Through such consistent adjustment, the alignment accuracy of the data can be improved and the reliability of the analysis results can be increased.

[0052] According to another embodiment, the system may further include an analysis software module, which is a software module that executes a high-resolution correction algorithm and a visualization UI. This analysis software module extends the capabilities of the ultra-precision temperature resolution-based digital melting curve analysis and visualization system to enable high-resolution correction and visual representation of data.

[0053] According to another embodiment, the analysis software module may include a data smoothing module that processes data smoothly to improve analysis accuracy. This data smoothing module serves to remove noise from data acquired in an ultra-precision temperature resolution-based digital melting curve analysis and visualization system and to improve the quality of the data.

[0054] According to another embodiment, the analysis software module may include a noise removal module that improves data quality by removing noise from the data. This noise removal module plays a role in enhancing the accuracy and reliability of data in a digital melting curve analysis and visualization system based on ultra-precision temperature resolution.

[0055] According to another embodiment, the analysis software module may include a threshold filter module that filters data based on a set threshold. This threshold filter module can remove unnecessary data during the analysis process or select and process only data that satisfies specific conditions. Through threshold filtering, the quality of the data can be improved and the accuracy of the analysis results can be increased.

[0056] According to the present invention, the accuracy and reliability of molecular biological experiments can be improved by analyzing and visualizing digital melting curves based on ultra-precision temperature resolution. Furthermore, by processing and analyzing data in real time, experimental time can be shortened and efficiency increased, and users can easily understand and interpret experimental results through an intuitive visualization interface.

[0057] The ultra-precision temperature resolution-based digital melting curve analysis and visualization system according to the present invention includes a data input module that uses raw melting data collected from a real-time digital PCR platform as input values. Additionally, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system includes a temperature resolution correction module that converts existing measured temperature intervals into a high-resolution curve. The ultra-precision temperature resolution-based digital melting curve analysis and visualization system includes a high-resolution Tm data processing module that precisely extracts interpolated Tm data.

[0058] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data input module (100), a temperature resolution correction module (200), and a high-resolution Tm data processing module (300). Preferably, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution (10) may be a system for analyzing and visualizing the melting characteristics of nucleic acids with high precision in digital PCR technology. Preferably, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution (10) may provide more precise analysis results by utilizing data obtained from real-time PCR equipment. Preferably, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution (10) may be implemented as a software module to overcome the hardware limitations of existing equipment through software.

[0059] Preferably, the data input module (100) can use raw melting data collected from a real-time digital PCR platform as input values. Preferably, the data input module (100) supports data files of various formats to process data generated from different PCR instruments in a compatible manner. Preferably, the data input module (100) can process a data set consisting of pairs of temperature and fluorescence intensity values. Preferably, the data input module (100) includes a data verification function to filter out incomplete or erroneous data. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include the data input module (100). Preferably, the temperature resolution correction module (200) can convert existing measured temperature intervals into high-resolution curves. Preferably, the temperature resolution correction module (200) can precisely estimate values ​​between temperature data points using linear interpolation, spline interpolation, polynomial interpolation, or interpolation based on Gaussian process regression. Preferably, the temperature resolution correction module (200) can convert temperature intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C, which are the measurement temperatures of a typical PCR instrument, into high resolution temperature intervals of 0.1°C, 0.05°C, or 0.01°C. Preferably, the temperature resolution correction module (200) may include a smoothing algorithm to minimize noise that may occur during the interpolation process. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include the temperature resolution correction module (200).

[0060] Preferably, the data input module (100) and the temperature resolution correction module (200) are linked in real time to immediately convert the input raw data into high-resolution data. Preferably, the temperature resolution correction module (200) and the high-resolution Tm data processing module (300) form a continuous data processing pipeline to analyze the interpolated data without delay. Preferably, all three modules support serial processing functions to efficiently process large datasets.

[0061] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may be implemented on a cloud basis, enabling data analysis through remote access. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may be developed as a cross-platform solution that operates on various operating systems. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may provide a user-friendly interface, allowing even non-expert users to easily utilize it.

[0062] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a real-time digital PCR platform interface module (110).

[0063] Preferably, the real-time digital PCR platform interface module (110) can collect data by linking with the real-time digital PCR platform. Preferably, the real-time digital PCR platform interface module (110) can be compatible with PCR equipment from various manufacturers through a standardized API (Application Programming Interface). Preferably, the real-time digital PCR platform interface module (110) can be connected to PCR equipment via a wired connection by supporting various communication protocols such as USB, Ethernet, Bluetooth, and Wi-Fi. Preferably, the real-time digital PCR platform interface module (110) can collect fluorescence signal data according to temperature changes in real time with an ultra-precision temperature resolution of 0.2°C or more, such as 0.2°C, 0.3°C, 0.5°C, or 1.0°C. Preferably, the data input module (100) can use raw melting data collected from the real-time digital PCR platform as input values. Preferably, the data input module (100) may include the real-time digital PCR platform interface module (110). Preferably, the data input module (100) supports data files of various formats (CSV, XML, JSON, ODPI, etc.) so that it can also analyze previously stored PCR data. Preferably, the data input module (100) evaluates the quality of the input data through a data integrity verification algorithm and can filter out data that is noisy or incomplete.

[0064] Preferably, the data input module (100) may have a parallel processing function capable of simultaneously processing fluorescence data from multiple channels generated in a multiplex PCR experiment. Preferably, the data input module (100) may also collect experiment metadata (experiment conditions, sample information, primer information, etc.) and use it for the interpretation of analysis results.

[0065] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a temperature resolution correction module (200) and an interpolation processing module (210).

[0066] Preferably, the temperature resolution correction module (200) can convert existing measured temperature intervals into a high-resolution curve. Preferably, the temperature resolution correction module (200) can improve the resolution of the temperature data measured in the melting curve analysis to enable more precise analysis. Preferably, the temperature resolution correction module (200) can increase the precision of the temperature data by applying various algorithms. Preferably, the temperature resolution correction module (200) may include an interpolation processing module (210).

[0067] Preferably, the interpolation processing module (210) can convert existing measurement data of temperature intervals of 0.2°C or more, e.g. 0.2°C, 0.3°C, 0.5°C, 1.0°C, into a high-resolution curve of 0.1°C or less by generating additional temperature points using a linear interpolation method. Preferably, the interpolation processing module (210) can calculate an intermediate value by assuming a linear relationship between two adjacent measurement data points. In one embodiment, the interpolation processing module (210) can generate additional data points at 0.1°C intervals between two points measured at an interval of 0.2°C or more, thereby adding a total of three new data points for each interval. In one embodiment, the interpolation processing module (210) can also generate ultra-high-resolution data points of 0.1°C, 0.05°C, or 0.01°C when higher precision is required. In one embodiment, the interpolation processing module (210) may perform interpolation after filtering noise from the original measurement data to increase the reliability of the interpolated data points. In one embodiment, the temperature resolution correction module (200) may include the interpolation processing module (210). Preferably, the high-resolution data generated by the interpolation processing module (210) can more accurately capture minute changes in the melting curve to improve the accuracy of the analysis. In one embodiment, the interpolation processing module (210) can process the data in real time to provide a high-resolution melting curve without delay. Preferably, the interpolation processing module (210) can operate stably under various experimental conditions to provide consistent results.Preferably, the interpolation processing module (210) may have various implementation examples, and in some implementation examples, in addition to linear interpolation, advanced interpolation techniques such as spline interpolation, polynomial interpolation, or Gaussian process regression-based interpolation may be optionally applied. Preferably, the interpolation processing module (210) may provide an interface that allows the user to set the desired temperature resolution. Preferably, the interpolation processing module (210) may display the interpolated data and the original data separately so that the user can clearly recognize the source of the data.

[0068] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a temperature resolution correction module (200) and a derivative curve generation module (220). Preferably, the temperature resolution correction module (200) can convert existing measured temperature intervals into a high-resolution curve. Preferably, the temperature resolution correction module (200) can receive and process fluorescence signals and temperature data measured in real time. Preferably, the temperature resolution correction module (200) can improve temperature resolution by subdividing the intervals between measured temperature points into smaller intervals through interpolation. Preferably, the temperature resolution correction module (200) can generate a high-resolution melting curve by utilizing mathematical algorithms such as linear interpolation, spline interpolation, polynomial interpolation, or interpolation based on Gaussian process regression. Preferably, the temperature resolution correction module (200) may include a derivative curve generation module (220).

[0069] Preferably, the derivative curve generation module (220) can extract the accurate Tm value of each well based on the first derivative (-dF / dT) curve of each well. Preferably, the derivative curve generation module (220) can adjust the resolution and smoothing degree of the derivative curve according to parameters specified by the user. Preferably, the derivative curve generation module (220) can automatically calculate characteristics such as the height, width, and area of ​​the peak in the generated derivative curve to provide additional analysis information.

[0070] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a high-resolution Tm data processing module (300) and a Tm data segmentation processing module (310).

[0071] Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) can process Tm data of nucleic acid molecules, such as DNA or RNA, to analyze the characteristics of the molecules. Preferably, the high-resolution Tm data processing module (300) can receive and process raw data obtained from a real-time PCR device or a digital PCR device. Preferably, the high-resolution Tm data processing module (300) can analyze the melting characteristics of nucleic acid molecules by measuring changes in fluorescence signals according to temperature. Preferably, the high-resolution Tm data processing module (300) may include a Tm data segmentation processing module (310). Preferably, the Tm data segmentation processing module (310) can generate a histogram by segmenting the x-axis in units of 0.1°C. Preferably, the Tm data segmentation processing module (310) can divide the Tm data into segments at intervals of 0.1°C and calculate the frequency of data corresponding to each segment. Preferably, the Tm data segmentation processing module (310) can provide a temperature resolution that is 2 to 100 times more precise than the existing resolution of 0.2°C, 0.3°C, 0.5°C, or 1°C units. Preferably, the Tm data segmentation processing module (310) can visually represent minute changes in Tm through the generated histogram. Preferably, the Tm data segmentation processing module (310) can provide a basis for precise analysis to distinguish the melting characteristics of various nucleic acid molecules. Preferably, the Tm data segmentation processing module (310) can identify the distribution characteristics of Tm by performing statistical analysis based on the segmented data.

[0072] Preferably, the high-resolution Tm data processing module (300) can generate a melting curve by additionally processing the histogram data generated by the Tm data segmentation processing module (310). Preferably, the high-resolution Tm data processing module (300) can determine the structural characteristics of nucleic acid molecules or whether there is a mutation through the generated melting curve. Preferably, the high-resolution Tm data processing module (300) can detect minute Tm differences that were difficult to distinguish with existing technology based on a precise temperature resolution of 0.1°C.

[0073] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a well count calculation module (320).

[0074] Preferably, the well count calculation module (320) can display the number of wells with the corresponding Tm on the y-axis. Preferably, the well count calculation module (320) can accurately calculate the number of wells with the corresponding Tm value for each temperature range based on interpolated Tm data. Preferably, the well count calculation module (320) can visualize the calculated number of wells in the form of a histogram so that the user can intuitively understand the distribution of a specific Tm value.

[0075] Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) may include a corresponding well number calculation module (320). Preferably, the high-resolution Tm data processing module (300) can derive an accurate Tm value by processing raw data measured with ultra-precision temperature resolution.

[0076] Preferably, the well count calculation module (320) can analyze the distribution of Tm values ​​within a temperature range specified by the user. Preferably, the well count calculation module (320) can calculate the well count with ultra-precision temperature resolution in units of 0.1℃, 0.05℃, or 0.01℃ to provide an accurate distribution even with minute temperature differences. Preferably, the well count calculation module (320) can analyze the calculated data using various statistical methods to provide statistical quantities such as the mean, median, and standard deviation.

[0077] Preferably, the well count calculation module (320) provides a function to compare and analyze multiple experimental data, allowing for a visual comparison of differences in Tm distribution under various experimental conditions. Preferably, the well count calculation module (320) can apply an outlier detection algorithm to identify wells with abnormal Tm values ​​and notify the user. Preferably, the well count calculation module (320) can provide a function to export the calculated well count data in various formats such as CSV, Excel, JSON, and ODPI, enabling further analysis.

[0078] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a high-resolution visualization module (330). Preferably, the high-resolution visualization module (330) can visualize Tm distributions that are close to each other with a resolution 3 to 5 times higher than the existing resolution, so as to be clearly distinguishable. Preferably, the high-resolution visualization module (330) can map Tm data in pixel units and convert it into a high-resolution graphic form. Preferably, the high-resolution visualization module (330) can express even minute temperature differences of 0.1°C or less, such as 0.1°C, 0.05°C, or 0.01°C, through a color gradient that is visually distinguishable. Preferably, the high-resolution visualization module (330) can provide a zoom-in / zoom-out function so that the user can zoom in on a specific temperature range to enable more detailed analysis. Preferably, the high-resolution visualization module (330) can enable the user to analyze data from various angles through various visualization options (heatmap, 3D surface plot, contour map, etc.).

[0079] Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) may include a high-resolution visualization module (330) that visualizes Tm distributions that are close to each other with a resolution 3 to 5 times higher than the existing resolution, so as to be clearly distinguishable. Preferably, the high-resolution Tm data processing module (300) can precisely estimate values ​​between data points by applying cubic spline interpolation to temperature data collected in real time. Preferably, the high-resolution Tm data processing module (300) can remove irregularities in temperature data that may occur during the measurement process through a noise filtering algorithm.

[0080] Preferably, the high-resolution visualization module (330) can simultaneously visualize the Tm distributions of multiple samples through a multi-sample comparison function, allowing for an intuitive understanding of the differences. Preferably, the high-resolution visualization module (330) can provide a function to export the visualized data in various formats such as PNG, SVG, and PDF, enabling its use in writing research reports or papers. Preferably, the high-resolution visualization module (330) can support a user-defined color palette to enable visual emphasis on specific temperature ranges.

[0081] Preferably, the high-resolution visualization module (330) can be accessed remotely through an interface, allowing for data analysis and visualization even outside the laboratory. Preferably, the high-resolution visualization module (330) can provide a tool to verify the reproducibility of experimental results through a comparative analysis function with past data.

[0082] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a PCR control peak reference alignment module (340). Preferably, the PCR control peak reference alignment module (340) may set the PCR control peak used in the melting analysis as a reference value and reference calibrate the Tm value of the entire sample based on this. Preferably, the PCR control peak reference alignment module (340) may provide accurate analysis results by correcting for minute temperature deviations that may occur for each sample. Preferably, the PCR control peak reference alignment module (340) may improve the accuracy of the system by using a PCR product of a known sequence as an internal standard.

[0083] Preferably, the PCR control peak reference alignment module (340) can utilize the Tm value of a standard control included in each experimental batch as a reference point to obtain consistent results even under various experimental conditions. Preferably, the PCR control peak reference alignment module (340) can increase data reproducibility by minimizing systematic errors that may occur between experiments. Preferably, the PCR control peak reference alignment module (340) can include an algorithm that can accurately identify and analyze each peak even in complex samples containing multiple PCR products. Preferably, the PCR control peak reference alignment module (340) can facilitate comparative analysis between samples by calculating the relative positional relationship between the reference peak and the peak to be analyzed. Preferably, the high-resolution Tm data processing module (300) can include the PCR control peak reference alignment module (340). Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) can perform more precise analysis based on the data aligned in the PCR control peak reference alignment module (340). Preferably, the PCR control peak reference alignment module (340) can provide flexibility to adjust the alignment method according to specific criteria defined by the user.

[0084] Preferably, the PCR control peak reference alignment module (340) can have versatility applicable to various types of melting curve data. Preferably, the PCR control peak reference alignment module (340) can be utilized in various application fields such as single nucleotide polymorphism (SNP) analysis, methylation analysis, and gene mutation detection.

[0085] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a PCR control peak reference alignment module (340).

[0086] Preferably, the PCR control peak reference alignment module (340) can consistently adjust the relative position relative to the reference point when comparing data before and after alignment. Preferably, the PCR control peak reference alignment module (340) can identify a specific reference point in the melting curve data and align the data based on it. Preferably, the PCR control peak reference alignment module (340) can convert melting curve data obtained from various samples into a normalized form to enable accurate comparative analysis. Preferably, the PCR control peak reference alignment module (340) can set the point where the rate of change of the fluorescence signal due to temperature change is maximum as the reference point. Preferably, this reference point may correspond to the Tm of a specific DNA sequence, thereby enabling consistent data comparison even under various experimental conditions.

[0087] Preferably, the PCR control peak reference alignment module (340) can adjust the melting curves of all samples by shifting the data around a reference point so that they have the same reference point. Preferably, this alignment process can correct deviations caused by minute temperature differences or equipment errors between experiments. Preferably, the PCR control peak reference alignment module (340) can adjust only the position while maintaining the original form and characteristics of the data during the alignment process. Preferably, this enables accurate comparative analysis without distortion of the data.

[0088] Preferably, the PCR control peak reference alignment module (340) can visualize the aligned data and provide it in the form of a graph that allows the user to intuitively compare. Preferably, through this visualization function, the researcher can easily identify subtle differences between various samples. Preferably, the PCR control peak reference alignment module (340) can provide various algorithms for setting reference points so that the user can select an alignment method suitable for the experimental purpose. Preferably, these algorithms may include various methods such as maximum peak-based, specific temperature-based, or specific fluorescence intensity-based.

[0089] Preferably, the PCR control peak-based alignment module (340) provides a function to compare data before and after alignment, allowing the user to directly verify the alignment effect. Preferably, this can increase the transparency and reliability of the alignment process.

[0090] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data correction module (400).

[0091] Preferably, the data correction module (400) may be a software module that executes a high-resolution correction algorithm and a visualization UI. Preferably, the data correction module (400) can collect and process ultra-precision temperature data to detect minute changes in the melting curve. Preferably, the high-resolution correction algorithm included in the data correction module (400) can achieve a temperature resolution of 0.1°C or less by minimizing noise and errors that may occur during the temperature measurement process. Preferably, the data correction module (400) can accurately identify the inflection point of the melting curve by processing and analyzing data in real time.

[0092] Preferably, the visualization UI of the data correction module (400) can provide various graphic elements and interactive functions so that the user can intuitively understand and interpret the melting curve data. Preferably, the visualization UI can support various visualization methods such as 2D and 3D graphs, heat maps, and contour plots so that the user can analyze the data from multiple angles. Preferably, the data correction module (400) can provide functions that allow the user to zoom in on a specific area of ​​interest or apply data filtering. Preferably, the data correction module (400) can provide a comparative analysis function with past data so that the user can identify the trend of change over time.

[0093] Preferably, the data correction module (400) can facilitate collaboration among multiple researchers or institutions by including cloud-based data storage and sharing functions. Preferably, the data correction module (400) can maintain the security of sensitive research data through data encryption and access control functions. Preferably, the data correction module (400) can support the documentation of research results by automatically generating reports in various formats.

[0094] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system may include a data correction module (400). Preferably, the data correction module (400) may have cross-platform compatibility so that it can be run on various operating systems. Preferably, the data correction module (400) may be designed so that even non-expert users can easily operate it through a user-friendly interface.

[0095] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data correction module (400) and a data smoothing module (410).

[0096] Preferably, the data correction module (400) may be a software module that executes a high-resolution correction algorithm and a visualization UI. Preferably, the data correction module (400) may collect and process melting curve data to provide an intuitive visualization interface to the user. Preferably, the data correction module (400) may receive temperature data in real time and analyze it precisely to derive important thermodynamic properties such as melting points. Preferably, the data correction module (400) may integrate various analysis algorithms to accurately interpret complex thermodynamic phenomena.

[0097] Preferably, the data smoothing module (410) can improve analysis accuracy by smoothing the data. Preferably, the data smoothing module (410) can generate a clearer melting curve by removing noise and irregular fluctuations that occur during the temperature measurement process. Preferably, the data smoothing module (410) can refine the data by utilizing various mathematical algorithms such as the moving average method, Gaussian filtering, and spline interpolation.

[0098] Preferably, the data correction module (400) may include a data smoothing module (410) that processes the data smoothly to improve analysis accuracy. Preferably, the data smoothing module (410) may operate as a preprocessing step within the data correction module (400) to improve the accuracy of subsequent analysis. Preferably, the data smoothing module (410) may apply a smoothing algorithm to incoming data in real time and automatically select an optimal smoothing method according to the characteristics of the data.

[0099] Preferably, the data smoothing module (410) can take a balanced approach that preserves important features of the raw data while eliminating unnecessary variations.

[0100] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a noise removal module (420). Preferably, the noise removal module (420) can improve data quality by removing noise from the data. Preferably, the noise removal module (420) may include an algorithm that identifies and removes various types of noise occurring in the digital melting curve data. Preferably, the noise removal module (420) can improve data quality by utilizing advanced signal processing techniques such as frequency domain filtering, wavelet transform, and Kalman filtering. Preferably, the noise removal module (420) can effectively respond to changing noise patterns by applying a noise removal algorithm to data input in real time. Preferably, the noise removal module (420) can selectively remove noise caused by various factors, such as thermal noise, electromagnetic interference, and sensor instability occurring during the temperature measurement process. Preferably, the noise removal module (420) can optimize the signal-to-noise ratio (SNR) to enable accurate detection of even minute temperature changes. Preferably, the data correction module (400) may include the noise removal module (420). Preferably, the data correction module (400) is a software module that executes a high-resolution correction algorithm and a visualization UI, and can improve data quality by organically linking with the noise removal module (420). Preferably, the data correction module (400) can derive accurate melting curve analysis results based on the clean data processed by the noise removal module (420). Preferably, the noise removal module (420) may include an intelligent system capable of automatically selecting and applying various noise removal algorithms according to data characteristics.

[0101] Preferably, the noise removal module (420) can implement a sophisticated algorithm that selectively removes only noise while preserving important characteristics of the data. Preferably, the noise removal module (420) can remove only unnecessary signal fluctuations while preserving fine inflection points or singularities of the melting curve.

[0102] An embodiment of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data correction module (400) and a threshold filter module (430). Preferably, the data correction module (400) may be a software module that executes a high-resolution correction algorithm and a visualization UI. Preferably, the data correction module (400) may process and analyze melting curve data to provide intuitive visualization results to the user. Preferably, the data correction module (400) may include various sub-modules to perform data collection, processing, analysis, and visualization functions. Preferably, the data correction module (400) may include a threshold filter module (430).

[0103] Preferably, the threshold filter module (430) can filter data based on a set threshold. Preferably, the threshold filter module (430) can remove noise or outliers from melting curve data based on a threshold defined by the user or a threshold automatically calculated by the system. Preferably, the threshold filter module (430) can improve the quality of the data by applying thresholds for the rate of temperature change, fluorescence signal intensity, or other related parameters. Preferably, the threshold filter module (430) can refine the data according to the user's analysis purpose by supporting various filtering algorithms. Preferably, the threshold filter module (430) can selectively extract only data within a specific range by simultaneously applying an upper Tm threshold and a lower Tm threshold. Preferably, the threshold filter module (430) can perform optimized filtering at various stages of the melting curve by applying dynamic thresholds that change over time. Preferably, the threshold filter module (430) can visualize the filtering results in real time so that the user can immediately verify the effect of the threshold setting.

[0104] Preferably, the data correction module (400) can perform advanced analysis functions based on the data filtered by the threshold filter module (430). Preferably, the threshold filter module (430) can store and manage filtering history so that the user can reuse or compare previously applied filtering settings. Preferably, the threshold filter module (430) can provide options to apply independent thresholds per channel or to apply a threshold integrated across all channels for multi-channel data. Preferably, the threshold filter module (430) can provide statistical summary information on data excluded during the filtering process so that the user can evaluate the impact of data loss.

[0106] Now, an embodiment of the present invention will be described in detail with reference to the attached drawings. Referring to FIG. 1, FIG. 1 is a conceptual block diagram of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution (10) may include a data input module (100), a temperature resolution correction module (200), and a high-resolution Tm data processing module (300). Preferably, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution (10) may be a system that analyzes and visualizes the melting characteristics of nucleic acids, such as DNA or RNA, at high resolution in digital PCR technology. Preferably, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution (10) may include a data input module (100). Preferably, the data input module (100) may use raw melting data collected from a real-time digital PCR platform as input values. Preferably, the data input module (100) can receive raw melting data of various formats and convert it into a normalized form to prepare it for subsequent analysis. Preferably, the data input module (100) can process raw melting data including fluorescence signal intensity data according to temperature changes. Preferably, the data input module (100) can automatically recognize data formats generated from various digital PCR instruments and extract data by applying an appropriate parsing algorithm. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include a temperature resolution correction module (200). Preferably, the temperature resolution correction module (200) can convert existing measured temperature intervals into high-resolution curves.

[0107] Preferably, the temperature resolution correction module (200) can precisely estimate values ​​between temperature data points using advanced mathematical algorithms such as linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression-based interpolation. Preferably, the temperature resolution correction module (200) can convert existing melting curve data, which is typically measured at intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C, into ultra-precision temperature resolution in units of 0.1°C or less, e.g., 0.1°C, 0.05°C, or 0.01°C.

[0108] Preferably, the temperature resolution correction module (200) may include an algorithm for detecting and filtering noise or artifacts that may occur during the interpolation process. Preferably, the temperature resolution correction module (200) may provide an interface that allows the user to select a desired temperature resolution. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include a high-resolution Tm data processing module (300). Preferably, the high-resolution Tm data processing module (300) may precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) may accurately identify Tm by calculating the derivative value of the melting curve. Preferably, the high-resolution Tm data processing module (300) may identify and separate multiple Tm values ​​in a complex melting curve using a multi-peak detection algorithm. Preferably, the high-resolution Tm data processing module (300) may calculate the distribution, standard deviation, confidence interval, etc. of Tm values ​​through a statistical analysis tool. Preferably, the high-resolution Tm data processing module (300) can support the user in intuitively interpreting the results by expressing the processed data in various visualization forms (histograms, scatter plots, heat maps, etc.). Preferably, the three modules of the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can operate in conjunction with each other.

[0109] Preferably, raw data collected from the data input module (100) can be transmitted to the temperature resolution correction module (200) and converted into high-resolution data.

[0110] Preferably, the converted high-resolution data is transmitted to a high-resolution Tm data processing module (300) so that precise Tm value extraction and analysis can be performed.

[0111] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can be implemented as a cloud-based service and provide an interface that can be accessed through a web browser.

[0112] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide the ability to export the analysis results in various formats (CSV, Excel, PDF, etc.).

[0113] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can safely protect sensitive research data by including user permission management and data security functions.

[0114] Referring to FIG. 2, FIG. 2 is a conceptual block diagram of a data input module according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data input module (100) and a real-time digital PCR platform interface module (110).

[0115] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system may be a system for effectively analyzing and visualizing raw melting data collected from a real-time digital PCR platform. Preferably, the data input module (100) may use raw melting data collected from a real-time digital PCR platform as input values. The data input module (100) may accept and process raw melting data of various formats and convert them into a form that can be analyzed within the system. The data input module (100) may process raw melting data including fluorescence signal intensity data according to temperature changes.

[0116] According to one embodiment of the present invention, a data input module (100) may include a real-time digital PCR platform interface module (110). Preferably, the real-time digital PCR platform interface module (110) may collect data in conjunction with a real-time digital PCR platform. The real-time digital PCR platform interface module (110) may provide a standardized interface compatible with digital PCR equipment from various manufacturers. The real-time digital PCR platform interface module (110) may support various communication protocols such as USB, Ethernet, Bluetooth, or Wi-Fi to provide flexibility in data transmission.

[0117] According to one embodiment of the present invention, the real-time digital PCR platform interface module (110) may include a function for filtering noise or errors that may occur during the data collection process. The real-time digital PCR platform interface module (110) may verify the integrity of the collected data and, if necessary, automatically perform correction operations. Preferably, the data input module (100) may include a function for storing and managing the collected raw melting data in an internal system database. The data input module (100) may provide an interface that allows the user to easily search for and reanalyze previously collected data. Preferably, the data input module (100) may manage metadata (experimental conditions, sample information, date, etc.) for the collected data together to ensure the traceability and reproducibility of the data. The data input module (100) may support data files of various formats (CSV, XML, JSON, ODPI, etc.) to enhance user convenience.

[0118] Referring to FIG. 3, FIG. 3 is a conceptual block diagram of a temperature resolution correction module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a temperature resolution correction module (200) and an interpolation processing module (210). Preferably, the temperature resolution correction module (200) can convert existing measured temperature intervals into a high-resolution curve. Preferably, the temperature resolution correction module (200) may be a key component for improving the precision of temperature data in digital melting curve analysis.

[0119] Preferably, the temperature resolution correction module (200) can receive and process temperature data measured at a low resolution.

[0120] Preferably, the temperature resolution correction module (200) can perform the function of analyzing the intervals of the input temperature data and converting them into data with denser intervals. Preferably, the temperature resolution correction module (200) can generate a more precise melting curve based on the converted high-resolution temperature data.

[0121] Preferably, the temperature resolution correction module (200) may include an interpolation processing module (210). Preferably, the interpolation processing module (210) can convert existing measurement data at intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C into a high-resolution curve in a temperature unit of 0.1°C or less by generating additional temperature points using a linear interpolation method. Preferably, the interpolation processing module (210) can generate a new data point by mathematically calculating the value between two adjacent temperature measurement points.

[0122] Preferably, the interpolation processing module (210) can assume a linear relationship between two adjacent measurement points and calculate values ​​on that line to perform interpolation. In one embodiment, the interpolation processing module (210) can generate two additional data points at a 0.1°C interval between two points measured at an interval of 0.2°C or more. Preferably, the interpolation processing module (210) can also generate data points at a finer interval, such as 0.05°C or 0.01°C, as needed.

[0123] Preferably, the interpolation processing module (210) may include an error analysis function to verify the accuracy of the interpolated data. Preferably, the interpolation processing module (210) may apply an algorithm to minimize errors that may occur during the interpolation process. Preferably, the interpolation processing module (210) is responsible for the core function of the temperature resolution correction module (200) and can play an important role in improving the analysis accuracy of the entire system.

[0124] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system may include a temperature resolution correction module (200). Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system may provide more precise melting curve analysis results based on high-resolution data processed through the temperature resolution correction module (200).

[0125] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system can visualize high-resolution temperature data to enable users to more accurately understand the characteristics of the melting curve according to minute temperature changes. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system can derive more accurate analysis results from high-resolution data by applying various analysis algorithms.

[0126] Referring to FIG. 4, FIG. 4 is a conceptual block diagram of a temperature resolution correction module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a temperature resolution correction module (200) and a derivative curve generation module (220).

[0127] Preferably, the temperature resolution correction module (200) can convert existing measured temperature intervals into a high-resolution curve. Preferably, the temperature resolution correction module (200) can receive and process real-time measured fluorescence signals and temperature data. Preferably, the temperature resolution correction module (200) can interpolate temperature data measured in general PCR equipment at intervals of 0.2°C to 1°C (0.2°C, 0.3°C, 0.5°C, 1°C) into ultra-precision temperature intervals of 0.1°C or less, for example, 0.05°C or 0.01°C. Preferably, the temperature resolution correction module (200) can precisely model the relationship between temperature and fluorescence signals by utilizing spline interpolation or polynomial interpolation.

[0128] Preferably, according to one embodiment of the present invention, the temperature resolution correction module (200) may include a derivative curve generation module (220). Preferably, the derivative curve generation module (220) can improve the precision of Tm extraction by smoothly generating the first derivative (-dF / dT) curve of each well. Preferably, the derivative curve generation module (220) can calculate the rate of change of the fluorescence signal by performing a differentiation operation based on the corrected high-resolution temperature-fluorescence data. Preferably, the derivative curve generation module (220) can improve the quality of the derivative curve by applying a smoothing algorithm to minimize noise.

[0129] Preferably, the derivative curve generation module (220) can automatically optimize various smoothing parameters to generate an optimal derivative curve suitable for each experimental condition. Preferably, the derivative curve generation module (220) can automatically analyze characteristics such as the position, height, and area of ​​peaks in the generated derivative curve to accurately determine the Tm value. Preferably, the derivative curve generation module (220) can accurately identify and separate each peak even in a complex melting curve containing multiple peaks.

[0130] Preferably, the temperature resolution correction module (200) and the derivative curve generation module (220) can operate in conjunction with each other. Preferably, high-resolution data generated by the temperature resolution correction module (200) is provided as input to the derivative curve generation module (220) to derive more precise analysis results. Preferably, data exchange between the two modules is performed in real time to provide immediate analysis results to the user.

[0131] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system can be designed to be compatible with various PCR instruments and experimental protocols. Preferably, the system can simplify complex analysis processes through a user-friendly interface and support researchers in easily interpreting results.

[0132] Referring to FIG. 5, FIG. 5 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a high-resolution Tm data processing module (300) and a Tm data segmentation processing module (310).

[0133] Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) can receive and process raw Tm data measured in real time. Preferably, the high-resolution Tm data processing module (300) can generate a continuous data flow by interpolating values ​​between measured data points through a mathematical algorithm. Preferably, the high-resolution Tm data processing module (300) can improve the precision of the data by using methods such as linear interpolation, spline interpolation, polynomial interpolation, or interpolation based on Gaussian process regression. Preferably, the high-resolution Tm data processing module (300) can remove unnecessary signals that may occur during the measurement process by applying a noise filtering algorithm.

[0134] Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing module (300) may include a Tm data segmentation processing module (310). Preferably, the Tm data segmentation processing module (310) may generate a histogram by segmenting the x-axis in units of 0.1°C. Preferably, the Tm data segmentation processing module (310) may classify interpolated Tm data into temperature segments with intervals of 0.1°C and calculate the frequency of data corresponding to each segment. Preferably, the Tm data segmentation processing module (310) may generate a visual representation in the form of a histogram based on the calculated frequency data. Preferably, the Tm data segmentation processing module (310) may be configured so that the size of the segments can be set to 0.01°C, 0.05°C, or 0.1°C according to user settings. Preferably, the high-resolution Tm data processing module (300) and the Tm data segmentation processing module (310) may interact with each other to provide precise melting curve analysis results. Preferably, data processed by the high-resolution Tm data processing module (300) is transmitted to the Tm data segmentation processing module (310) and can be visualized through segmentation processing. Preferably, through this processing, minute changes in the melting curve can be detected with a temperature resolution improved by up to 100 times compared to existing systems. Preferably, the high-resolution Tm data processing module (300) can support the analysis of various types of biological samples. Preferably, the high-resolution Tm data processing module (300) may include an algorithm capable of analyzing the melting characteristics of various biomolecules such as DNA, RNA, and proteins. Preferably, the high-resolution Tm data processing module (300) can maintain data consistency in various environments by providing a correction function according to experimental conditions.

[0135] Preferably, the Tm data interval processing module (310) can provide a function to store the generated histogram data and compare and analyze it with previous experimental results. Preferably, the Tm data interval processing module (310) can facilitate visual comparison by displaying the histograms of multiple samples in a superimposed manner. Preferably, the Tm data interval processing module (310) can quantify characteristics such as the peaks, distribution width, and symmetry of the histogram through a statistical analysis tool.

[0136] Referring to FIG. 6, FIG. 6 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a high-resolution Tm data processing module (300) and a well count calculation module (320). Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) can receive and process raw Tm data collected in real time. Preferably, the high-resolution Tm data processing module (300) can perform a filtering process to remove noise from the collected Tm data and improve data quality. Preferably, the high-resolution Tm data processing module (300) can precisely estimate values ​​between measured temperature points by utilizing linear interpolation, spline interpolation, polynomial interpolation, or interpolation based on Gaussian process regression. Preferably, the high-resolution Tm data processing module (300) can detect even minute temperature changes by processing Tm data with a temperature resolution of 0.1°C or less, such as 0.1°C, 0.05°C, or 0.01°C. Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing module (300) may include a well count calculation module (320). Preferably, the well count calculation module (320) can represent the number of wells with the corresponding Tm on the y-axis. Preferably, the well count calculation module (320) can accurately calculate the number of wells with the corresponding Tm for each temperature interval. Preferably, the well count calculation module (320) can process the calculated well count data by converting it into a histogram form to facilitate visual analysis.Preferably, the well count calculation module (320) can adjust the resolution of the well count calculation according to the temperature range specified by the user.

[0137] Preferably, the high-resolution Tm data processing module (300) and the well count calculation module (320) can be interconnected to enable precise melting curve analysis. Preferably, the precise Tm data processed by the high-resolution Tm data processing module (300) can be input into the well count calculation module (320) and used to calculate the well distribution for each temperature. Preferably, data exchange between the two modules can be performed in real time to minimize delays in the analysis process. Preferably, the high-resolution Tm data processing module (300) can provide an interface that allows parameters to be adjusted according to various experimental conditions. Preferably, the well count calculation module (320) can include a function to export the calculated data in various formats (CSV, Excel, JSON, ODPI, etc.). Preferably, the two modules work together to detect minute changes in the melting curve, thereby enabling more accurate analysis of the characteristics of DNA or RNA samples.

[0138] Referring to FIG. 7, FIG. 7 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a high-resolution Tm data processing module (300) and a high-resolution visualization module (330).

[0139] Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) can receive and process raw Tm data collected in real time. Preferably, the high-resolution Tm data processing module (300) can perform noise removal and signal normalization processes on the collected raw data. Preferably, the high-resolution Tm data processing module (300) can precisely estimate values ​​between data points by utilizing advanced mathematical algorithms such as linear interpolation, spline interpolation, polynomial interpolation, or interpolation based on Gaussian process regression. Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing module (300) may include a high-resolution visualization module (330). Preferably, the high-resolution visualization module (330) can visualize Tm distributions that are close to each other with a resolution 3 to 5 times higher than the existing resolution, so as to be clearly distinguishable. Preferably, the high-resolution visualization module (330) can receive and process precise Tm data extracted from the high-resolution Tm data processing module (300). Preferably, the high-resolution visualization module (330) can utilize various color mapping techniques to visually distinguish even minute temperature differences. Preferably, the high-resolution visualization module (330) can provide a zoom function that allows the user to zoom in on a specific temperature range. Preferably, the high-resolution visualization module (330) can support various visualization methods, such as 3D graphs, heat maps, and contour graphs, to enable the user to analyze the data from multiple angles.Preferably, the high-resolution Tm data processing module (300) and the high-resolution visualization module (330) can operate in close coordination with each other. Preferably, data processed by the high-resolution Tm data processing module (300) can be immediately transmitted to the high-resolution visualization module (330) for visualization. Preferably, when a user selects a specific data area through the high-resolution visualization module (330), the high-resolution Tm data processing module (300) can perform a more detailed analysis of that area. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system may include an ultra-high sensitivity sensor system capable of detecting and analyzing temperature differences of 0.1°C or less. Preferably, this system can be utilized to analyze minute structural differences of biomolecules such as DNA, RNA, and proteins in fields such as molecular biology, genomics, and proteomics. Furthermore, this system can be effectively applied to various molecular diagnostic and precision medicine applications, such as SNP-based mutation analysis, precise epigenetic analysis at the single base methylation level, and copy number variation (CNV) detection.

[0140] Referring to FIG. 8, FIG. 8 is a conceptual block diagram of a high-resolution Tm data processing module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a high-resolution Tm data processing module (300) and a PCR control peak reference alignment module (340).

[0141] Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing module (300) can receive and process raw fluorescence data obtained from a real-time PCR device. Preferably, the high-resolution Tm data processing module (300) can monitor the dissociation process of DNA double strands by measuring changes in the fluorescence signal according to an increase in temperature. Preferably, the high-resolution Tm data processing module (300) can determine the Tm value by precisely identifying the temperature point where a rapid decrease in the fluorescence signal occurs. Preferably, the high-resolution Tm data processing module (300) can overcome the temperature resolution limitations of existing equipment by subdividing the intervals between temperature data points through interpolation. Preferably, the high-resolution Tm data processing module (300) can improve the resolution of temperature data by using methods such as linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression-based interpolation. Preferably, the high-resolution Tm data processing module (300) can more accurately identify Tm peaks by calculating the first derivative or second derivative curve of the fluorescence signal based on interpolated data. Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing module (300) may include a PCR control peak reference alignment module (340). Preferably, the PCR control peak reference alignment module (340) can set the PCR control peak used in the melting analysis as a reference value and reference calibrate the Tm value of the entire sample based on this.Preferably, the PCR control peak reference alignment module (340) can improve the comparability between experiments by correcting for minute temperature fluctuations that may occur in each experimental batch. Preferably, the PCR control peak reference alignment module (340) can set the PCR control peak value using control DNA of a known sequence. Preferably, the PCR control peak reference alignment module (340) can derive a normalized Tm value by calculating the relative difference between the PCR control peak value and the Tm value of each sample. Preferably, the PCR control peak reference alignment module (340) can provide consistent Tm data despite differences in various experimental conditions or equipment.

[0142] Preferably, the PCR control peak reference alignment module (340) can continuously monitor the stability of the PCR control peak value to detect outliers that may occur during the experiment. Preferably, the PCR control peak reference alignment module (340) can improve alignment accuracy in various temperature ranges by using multiple PCR control peaks.

[0143] Preferably, the high-resolution Tm data processing module (300) can transmit the processed data to a visualization module so that the user can intuitively interpret the results. Preferably, the high-resolution Tm data processing module (300) can provide a function to store the processed Tm data in conjunction with a database and to compare and analyze it with previous experimental results.

[0144] Referring to FIG. 9, FIG. 9 is a conceptual block diagram of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. According to an embodiment of the present invention, the digital melting curve analysis and visualization system based on ultra-precision temperature resolution (10) may include a data correction module (400).

[0145] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may be a system for analyzing and visualizing the melting curve of DNA or RNA of a biological sample.

[0146] Preferably, the data correction module (400) may be a software module that executes a high-resolution correction algorithm and a visualization UI. Preferably, the data correction module (400) may collect and process melting curve data to precisely analyze the double-strand dissociation process of DNA or RNA according to temperature changes.

[0147] Preferably, the data correction module (400) can process melting curve data with an ultra-precision temperature resolution of 0.1°C or less, such as 0.1°C, 0.05°C, or 0.01°C. Preferably, the data correction module (400) can minimize measurement errors and calculate accurate Tm through a temperature correction algorithm. Preferably, the data correction module (400) can detect even minute changes by generating first and second derivative graphs of the melting curve. Preferably, the data correction module (400) can intuitively represent complex melting curve data by providing a user-friendly visualization UI.

[0148] Preferably, the data correction module (400) may include a function to simultaneously compare and analyze the melting curves of multiple samples. Preferably, the data correction module (400) may provide a function to export the analysis results in various formats (CSV, PDF, image, etc.).

[0149] Preferably, the data correction module (400) can enable remote collaboration by supporting cloud-based data storage and sharing functions. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include the data correction module (400). Preferably, the data correction module (400) can be optimized for various experimental conditions by providing a user-defined analysis parameter setting function.

[0150] Preferably, the data correction module (400) may include a specialized algorithm for identifying various types of gene mutations (methylation, MSI, SNP, insertion, deletion, etc.).

[0151] Referring to FIG. 10, FIG. 10 is a conceptual block diagram of an analysis software module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data correction module (400) and a data smoothing module (410).

[0152] Preferably, the data correction module (400) may be a software module that executes a high-resolution correction algorithm and a visualization UI. Preferably, the data correction module (400) may function as a core component of a digital melting curve analysis and visualization system based on ultra-precision temperature resolution. Preferably, the data correction module (400) may perform the function of collecting, processing, and analyzing melting curve data. Preferably, the data correction module (400) may precisely measure and analyze changes in the physical state of a sample according to temperature changes. Preferably, the data correction module (400) may visually represent the analysis results through a user interface so that the user can easily understand them. Preferably, according to one embodiment of the present invention, the data correction module (400) may include a data smoothing module (410). Preferably, the data smoothing module (410) may improve analysis accuracy by smoothly processing the data. Preferably, the data smoothing module (410) may improve the quality of the data by removing noise or outliers that may occur in the melting curve data. Preferably, the data smoothing module (410) can process data by applying various smoothing algorithms, such as the moving average method, Gaussian filter, and Savitzky-Golay filter. Preferably, the data smoothing module (410) can provide a parameter setting function that allows the user to adjust the smoothing intensity and method. Preferably, the data correction module (400) and the data smoothing module (410) can interact with each other to automate a series of processes from the collection of melting curve data to analysis and visualization. Preferably, the data correction module (400) can calculate important parameters such as the melting point, crystallization temperature, and enthalpy change based on the data processed by the data smoothing module (410).Preferably, the data correction module (400) can visualize the calculated parameters in various forms, such as graphs, charts, and tables, and provide them to the user.

[0153] Preferably, the data correction module (400) can provide a function to compare and analyze data obtained under various experimental conditions. Preferably, the data correction module (400) can track changes in the characteristics of the sample through comparative analysis with past data. Preferably, the data smoothing module (410) can process data in real time and provide it to the data correction module (400). Preferably, the data smoothing module (410) can derive optimal results by applying a smoothing algorithm according to the characteristics of the data. Preferably, the data smoothing module (410) can provide a visualization function that allows comparison of data before and after processing, enabling the user to intuitively check the smoothing effect.

[0154] Referring to FIG. 11, FIG. 11 is a conceptual block diagram of an analysis software module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data correction module (400) and a noise removal module (420). Preferably, the data correction module (400) may be a software module that executes a high-resolution correction algorithm and a visualization UI. Preferably, the data correction module (400) is a core component of the digital melting curve analysis and visualization system based on ultra-precision temperature resolution and can generate a melting curve by processing and analyzing collected temperature data. Preferably, the data correction module (400) can detect even minute changes in temperature data to provide a high-resolution melting curve. Preferably, the data correction module (400) can visually represent the analysis results through a user-friendly interface. Preferably, according to one embodiment of the present invention, the data correction module (400) may include a noise removal module (420). Preferably, the noise removal module (420) can improve data quality by removing noise from the data. Preferably, the noise removal module (420) can implement an algorithm to identify and remove noise, such as random fluctuations, electrical interference, and sensor errors, that may occur in the collected temperature data. Preferably, the noise removal module (420) can improve data quality by utilizing advanced signal processing techniques such as Fourier transform, wavelet transform, and Kalman filtering. Preferably, the noise removal module (420) can provide a function to visually verify the noise removal effect by comparing the raw data with the processed data. Preferably, the noise removal module (420) can provide parameter setting options that allow the user to adjust the noise removal intensity.Preferably, the noise removal module (420) can process data in real time to provide high-quality melting curve analysis results without delay.

[0155] Preferably, the data correction module (400) and the noise removal module (420) can interact with each other to improve the accuracy and reliability of the ultra-precision temperature resolution-based melting curve analysis system. Preferably, the data correction module (400) can perform more accurate melting point identification and analysis based on the data processed by the noise removal module (420). Preferably, efficient data flow between these modules can optimize the performance of the entire system.

[0156] Referring to FIG. 12, FIG. 12 is a conceptual block diagram of an analysis software module according to an example of an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a data correction module (400) and a threshold filter module (430). Preferably, the data correction module (400) may be a software module that executes a high-resolution correction algorithm and a visualization UI. Preferably, the data correction module (400) may perform the function of processing and analyzing collected data as a core component of the digital melting curve analysis and visualization system based on ultra-precision temperature resolution. Preferably, the data correction module (400) may play the role of collecting melting curve data, converting it to high resolution, and visualizing it in a form that is easy for the user to understand. Preferably, the data correction module (400) may include various sub-modules to manage the entire process from data collection to analysis and visualization. Preferably, according to one embodiment of the present invention, the data correction module (400) may include a threshold filter module (430). Preferably, the threshold filter module (430) can perform the function of filtering data based on a set threshold. Preferably, the threshold filter module (430) can select data based on a threshold set by the user to remove noise or outliers from the collected melting curve data. Preferably, the threshold filter module (430) can provide the function of selectively extracting only data within a specific temperature range or filtering data above or below a specific fluorescence intensity. Preferably, the threshold filter module (430) can provide an interface that allows the user to directly set the threshold, or include an algorithm that can automatically calculate and apply the optimal threshold.Preferably, the threshold filter module (430) can visualize the filtered result in real time so that the user can immediately check the effect of the threshold adjustment.

[0157] Preferably, the threshold filter module (430) and the data correction module (400) can interact to derive more accurate melting curve analysis results based on the filtered data. Preferably, the data processed by the threshold filter module (430) can undergo additional analysis and visualization processes by other components of the data correction module (400).

[0158] Preferably, the data correction module (400) may include various functional modules, such as a data collection module, a temperature correction module, and a visualization module, in addition to the threshold filter module (430), to provide a comprehensive melting curve analysis environment. Preferably, these modules are organically connected to each other to facilitate the flow of data and provide the user with an intuitive analysis experience.

[0159] Referring to FIG. 13, FIG. 13 illustrates an example of a melting curve correction method according to an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may include a temperature resolution correction module (200) and an interpolation processing module (210). Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be an integrated system for precise analysis and visualization of melting curve data. Preferably, the temperature resolution correction module (200) may convert existing measured temperature intervals into a high-resolution curve. Preferably, the temperature resolution correction module (200) may receive and process existing melting curve data measured at low resolution.

[0160] Preferably, the temperature resolution correction module (200) can improve the precision of the melting curve by converting data with relatively wide intervals (e.g., 0.3°C intervals) in the existing measurement data into data with denser intervals.

[0161] Preferably, the temperature resolution correction module (200) can generate a continuous and precise melting curve based on interpolated high-resolution data.

[0162] Preferably, according to one embodiment of the present invention, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include a temperature resolution correction module (200). Preferably, the interpolation processing module (210) can convert existing measurement data at intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C into a high-resolution curve of 0.1°C or less by generating additional temperature points using a linear interpolation method. Preferably, the interpolation processing module (210) can mathematically calculate a new data point between two adjacent measurement data points by assuming a linear relationship between them. Preferably, the interpolation processing module (210) can subdivide intervals of 0.2°C or more into intervals of 0.1°C or less by generating two or more new data points between each point of the existing measurement data.

[0163] Preferably, the interpolation processing module (210) can connect interpolated data points to form a smoother and more continuous melting curve. Preferably, the interpolation processing module (210) can improve measurement precision by overcoming the physical limitations of existing hardware through software methods. Preferably, according to one embodiment of the present invention, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include the interpolation processing module (210).

[0164] Preferably, the temperature resolution correction module (200) and the interpolation processing module (210) can be coupled to more accurately extract the peak position (Tm) of the melting curve. Preferably, the melting curve converted to high resolution can be particularly useful in applications requiring the analysis of minute Tm differences, such as SNP typing and single methylation identification. Preferably, the system of the present invention can significantly improve the precision of melting curve analysis through a software-based interpolation algorithm without modifying existing hardware. Preferably, as shown in FIG. 13, the graph on the left represents low-resolution data (e.g., in units of 0.3°C) collected from an existing system, and the graph on the right represents high-resolution results (in units of 0.1°C) to which the interpolation algorithm according to the present invention has been applied. This allows for more accurate extraction of the peak position (Tm) of the curve and is particularly advantageous for SNP typing and single methylation identification, where minute Tm differences need to be analyzed.

[0165] Preferably, through the interpolation process, new data points are generated between existing data so that the melting curve can be transformed from a dotted line form into a continuous and precise curve form.

[0166] Referring to FIG. 14, FIG. 14 illustrates an example of a peak curve based on a first derivative (-dF / dT) according to an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may include a temperature resolution correction module (200) and a threshold filter module (430). Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be a system for analyzing and visualizing the melting curves of various biological samples. Preferably, the temperature resolution correction module (200) may convert existing measured temperature intervals into high-resolution curves. Preferably, the temperature resolution correction module (200) may process data measured at existing low resolution (e.g., 0.3°C units) by normalizing it into high-resolution temperature data at 0.1°C units. Preferably, the temperature resolution correction module (200) can calculate the fluorescence signal value of an unmeasured intermediate temperature point using interpolation. Preferably, the temperature resolution correction module (200) can generate high-resolution data using methods such as linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression-based interpolation.

[0167] Preferably, the temperature resolution correction module (200) is a core component of the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) and can distinguish minute Tm differences between samples.

[0168] Preferably, the threshold filter module (430) can filter data based on a set threshold.

[0169] Preferably, the threshold filter module (430) can select only peaks having a signal intensity greater than or equal to the threshold value based on the threshold value indicated by the red baseline (Threshold) in the drawing as valid analysis targets. Preferably, the threshold filter module (430) can improve the reliability of the data by removing signals caused by noise or non-specific responses.

[0170] Preferably, the threshold filter module (430) provides an interface that allows the user to adjust the threshold value, thereby enabling the setting of optimal filtering criteria according to experimental conditions or sample characteristics. Preferably, the threshold filter module (430) serves as a component of the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) and can contribute to increasing the accuracy and reproducibility of the analysis results. Preferably, the temperature resolution correction module (200) and the threshold filter module (430) can operate in conjunction with each other, and the threshold filter module (430) can be applied to the high-resolution data generated by the temperature resolution correction module (200) to select the final analysis target peak.

[0171] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can accurately distinguish and analyze the melting points of different samples having Tm values ​​that are close to each other, as indicated by the blue arrows in the drawing. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can clearly separate and represent samples with close Tm values ​​that might be mistaken for a single peak in conventional low-resolution systems. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide intuitive analysis results to the user by generating a graph that displays temperature (°C) on the x-axis and the slope of the fluorescence signal on the y-axis. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can enable accurate peak quantification through histogram-based peak separation technology.

[0172] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can be utilized in various biological applications, particularly DNA melting analysis, SNP (single nucleotide polymorphism) detection, or gene mutation analysis.

[0173] Referring to FIG. 15, FIG. 15 is a diagram illustrating the effect of a histogram-based high-resolution Tm analysis and normalization technique according to an embodiment of the present invention compared with a conventional method. According to an embodiment of the present invention, a digital melting curve analysis and visualization system based on ultra-precision temperature resolution may include a high-resolution Tm data processing module (300) and a Tm data segmentation processing module (310).

[0174] Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data.

[0175] Preferably, the high-resolution Tm data processing module (300) can process Tm data with a much more precise temperature resolution than the existing approximately 0.5°C unit resolution.

[0176] Preferably, the high-resolution Tm data processing module (300) can analyze raw data obtained from the melting curve of a nucleic acid sample, such as DNA or RNA, to precisely detect changes in the fluorescence signal according to minute temperature changes.

[0177] Preferably, the high-resolution Tm data processing module (300) can perform the function of normalizing the Tm values ​​of the entire sample based on the representative value of the PCR control (Tm reference). Preferably, the high-resolution Tm data processing module (300) can eliminate deviations according to experimental conditions to enable quantitative comparison between data obtained under various conditions.

[0178] Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing module (300) may include a Tm data segmentation processing module (310).

[0179] Preferably, the Tm data interval processing module (310) can generate a histogram by intervalizing the x-axis in units of 0.1℃.

[0180] Preferably, the Tm data segmentation processing module (310) can separate multiple targets having similar Tm, which were difficult to distinguish because they were combined into a single peak at the existing 0.2°C, 0.3°C, 0.5°C, or 1.0°C unit resolution, so that they can be visually clearly distinguished.

[0181] Preferably, the Tm data segmentation processing module (310) can improve the sensitivity of fine sequence differences, such as single methylation or SNPs, through precise binning processing.

[0182] Preferably, the Tm data segmentation processing module (310) can restore information that was overlooked in the existing melting system and enable precise methylation pattern recognition. Preferably, the Tm data segmentation processing module (310) can align to 87.0 °C based on the PCR control peak of each sample to compare Tm distributions while removing deviations due to experimental conditions. Preferably, the Tm data segmentation processing module (310) can clearly distinguish relative differences in target Tm by performing alignment based on the PCR control peak.

[0183] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system of the present invention can provide an environment in which a user can quantitatively interpret the Tm distribution through a histogram UI. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system of the present invention can not only improve simple resolution but also structurally improve inter-experimental reproducibility and data consistency. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system of the present invention can play a key role in implementing a highly reliable methylation diagnostic platform when applied clinically.

[0184] Referring to FIG. 16, FIG. 16A is a diagram illustrating a comparative example of RT-qPCR-based multiplex melting curve analysis as background technology of the present invention. FIG. 16B is a diagram showing the results of digital PCR-based melting curve analysis according to an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may include a high-resolution Tm data processing module (300).

[0185] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can perform accurate identification and quantification of DNA templates through digital PCR-based melting curve analysis.

[0186] Preferably, the high-resolution Tm data processing module (300) can precisely extract interpolated Tm data.

[0187] Preferably, the high-resolution Tm data processing module (300) can achieve an ultra-precision temperature resolution of 0.1°C or less by applying an interpolation algorithm to raw melting curve data obtained based on digital PCR.

[0188] Preferably, the high-resolution Tm data processing module (300) can individually analyze the melting curve generated in each well to extract accurate Tm values ​​without interference between templates.

[0189] Preferably, the high-resolution Tm data processing module (300) can convert the extracted Tm data into a histogram form to visualize the distribution of each template.

[0190] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) may include a high-resolution Tm data processing module (300). Preferably, the high-resolution Tm data processing module (300) can clearly distinguish and analyze different DNA templates even if they have similar Tm values.

[0191] Preferably, the high-resolution Tm data processing module (300) can provide consistent Tm analysis results regardless of changes in the concentration of the template.

[0192] Preferably, the high-resolution Tm data processing module (300) can effectively resolve quantitative errors and Tm interference problems that occur in conventional RT-qPCR-based analysis.

[0193] Preferably, the high-resolution Tm data processing module (300) can preserve the unique melting curve pattern of each template even in a multiplex analysis environment to provide accurate analysis results.

[0194] Preferably, the high-resolution Tm data processing module (300) can provide excellent resolution and sensitivity even for the detection of small amounts of templates.

[0195] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide the same melting curve results whether four different DNA templates are used or a single template is used, as shown on the right side (B) of FIG. 16.

[0196] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can solve the problem of curve shape distortion due to the relative quantitative difference between templates and low temperature resolution, which is a limitation of the conventional RT-qPCR-based analysis shown on the left (A) of FIG. 16.

[0197] Preferably, the high-resolution Tm data processing module (300) can preserve the unique melting curve pattern for each template through analysis of each well unit, and process changes in template concentration so that only the absolute amount on the histogram changes and does not affect the Tm position.

[0198] Referring to FIG. 17, FIG. 17 is a conceptual diagram illustrating a melting curve analysis design for a specific gene promoter according to an embodiment of the present invention. According to one embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be included.

[0199] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can analyze the melting curve when a single CpG methylation occurs in a CpG island within the promoter region of a specific gene.

[0200] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can implement a temperature step of less than 0.1 °C to provide a resolution 3 to 5 times better than conventional analysis.

[0201] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can visualize a single change within a sulfur dioxide-treated CpG island.

[0202] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can specifically identify the difference in Tm between the methylated variant or mutation template and the Wild Type (WT). Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can simultaneously perform not only simple presence or absence determination (qualitative analysis) but also quantitative analysis based on the degree of single CpG methylation through digital melting analysis technology. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can separate and interpret minute methylation pattern differences that were not observed in existing systems with high precision.

[0203] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can improve the Tm extraction precision and resolution compared to existing systems. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can predict the Tm change of each single methylated template with high resolution.

[0204] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can quantitatively distinguish changes in a single CpG methylation level through high-resolution melting analysis. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can be used for disease diagnosis, drug response prediction, or research on gene expression regulation mechanisms by analyzing methylation patterns occurring in CpG islands of specific gene promoter regions.

[0205] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can monitor and analyze the dissociation process of the DNA double helix structure in real time according to temperature changes.

[0206] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can detect the dissociation process of the DNA double helix through changes in fluorescence signals and convert it into a digital signal for analysis.

[0207] Referring to FIG. 18, FIG. 18 is a diagram illustrating primer locations and methylation target regions designed based on the CpG island sequence within the promoter of the CDH13 gene as a specific example of the analysis design illustrated in FIG. 17 according to an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be included. Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may perform methylation analysis using primers designed based on the CpG island sequence within the promoter of the CDH13 gene. Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be specially designed so that the primers for PCR amplification have a specific Tm value.

[0208] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can selectively set only the sites having a Cytosine-Guanine (CpG) structure as methylation targets. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can perform analysis by reflecting the biochemical characteristics that methylation hardly occurs in Cytosine-Cytosine, Cytosine-Thymine, and Cytosine-Adenine structures.

[0209] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can observe melting peaks separated according to the number of CpG methylations by performing digital melting curve analysis using designed primers. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can visually separate and represent patterns in which two or more CpGs are methylated sequentially, starting from the case where one CpG is methylated. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can identify methylated CpG sites and quantitatively analyze the degree of methylation (hypermethylation) through melting curve analysis. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can detect the presence of a single methylation event at high resolution through the Tm difference of the methylated variant template compared to the wild type (WT).

[0210] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide very high sensitivity in both quantitative interpretation and qualitative discrimination through digital melting analysis. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can detect even minute differences in Tm by providing ultra-precision temperature resolution of 0.1°C or less. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can monitor changes in methylation patterns by collecting and analyzing melting curve data in real time.

[0211] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide a customized primer design function for methylation analysis of promoter regions of various genes. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can display collected melting curve data through an intuitive visualization interface so that the user can easily interpret the methylation pattern.

[0212] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can identify biomarkers that can be used for disease diagnosis or prognosis prediction based on methylation analysis results.

[0213] Referring to FIG. 19, FIG. 19 is a diagram quantitatively showing the results of a digital melting analysis performed based on the design of FIG. 17 and FIG. 18 according to an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be included.

[0214] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can set a reference peak included in each sample as a reference value and analyze the degree of CpG methylation based on ΔTm (Tm difference) with the target peak.

[0215] Preferably, the position of the reference peak may vary slightly depending on the conditions, but it can be used as a relative standard to quantitatively calculate the temperature difference from the target Tm of each sample.

[0216] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can show the largest difference with ΔTm 10.2 °C in unmethylated.

[0217] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can show a ΔTm of 9.8 °C in 1-methylation. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can show a ΔTm of 9.3 °C in 2-methylation.

[0218] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can show a ΔTm of 8.7 °C in 3-methylation.

[0219] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can show a ΔTm of 8.3 °C in 4-methylation.

[0220] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can visually show that the temperature difference from the reference decreases as the target Tm increases with increasing methylation levels. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can confirm that even a single CpG methylation difference between unmethylation and 1-methylation is clearly separated on the histogram through high-resolution Tm analysis technology in units of 0.1 °C. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can precisely distinguish even minute Tm changes that were difficult to observe in conventional systems.

[0221] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can demonstrate that quantitative analysis of methylation levels is possible through reference-based correction and ΔTm calculation, beyond simple qualitative determination.

[0222] Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution can be utilized as a high-performance analysis tool that can simultaneously satisfy sensitivity and precision in a digital melting analysis platform.

[0223] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can precisely measure changes in Tm according to various methylation levels and quantitatively analyze methylation patterns.

[0224] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can accurately predict the degree of methylation through a gradual decrease pattern of ΔTm values ​​according to the methylation level. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can determine the methylation level of an unknown sample by database-ing characteristic ΔTm values ​​for each methylation step.

[0225] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can visualize the temperature difference between the reference peak and the target peak so that the user can intuitively understand the methylation level.

[0226] Referring to FIG. 20, FIG. 20 is a conceptual diagram illustrating the progression of methylation histogram analysis and abnormal DNA methylation according to an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be included. Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may generate a histogram by performing digital melting analysis on the CpG region of a healthy individual. Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may construct a histogram by displaying melting points (°C) on the horizontal axis and the number of cells having corresponding melting points on the vertical axis. Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may perform high-precision absolute quantitative analysis approaching the single-cell level through digital PCR-based analysis. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can construct a high-resolution histogram based on the number of cells having the same melting point. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can visualize the pathological progression of abnormal DNA methylation induced by external physical, chemical, or biological stimuli step by step.

[0227] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can represent abnormal DNA methylation occurring at a very small number of CpG sites in the initial stage with a narrow range of data dispersion.

[0228] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can analyze the process of methylation that has not been normalized by a recovery mechanism within the human body gradually accumulating within the same cell.

[0229] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can detect the phenomenon of gene expression suppression caused by the methylation of tumor suppressor genes.

[0230] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can analyze DNA damage cell formation due to DNA repair failure.

[0231] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can monitor the process of tumor initiation caused by the accumulation of damaged cells.

[0232] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can detect pathological metastasis to cancer that progresses to a malignant state due to a rapid increase in abnormal DNA methylation.

[0233] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can repeatedly perform digital melting analysis on the same sample over time to track and observe time-series changes in Tm distribution and methylation status.

[0234] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can continuously monitor changes in the number of copies of a mutant template relative to a wild type (Wild Type, WT) to identify the trend of cell damage or genetic abnormalities accumulating.

[0235] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can be utilized in the healthcare field to detect disease progression in advance or to evaluate treatment effects over the long term. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can build a time-series prediction model based on individual patient or group data and accumulate the relevant data into a structured database (DB).

[0236] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide an important technical foundation in terms of precision medicine, such as early cancer diagnosis and progression prediction tools as well as healthcare, through high-resolution-based tracking analysis.

[0237] Referring to FIG. 21, FIG. 21 conceptually illustrates a method for detecting microsatellite instability (MSI) based on digital melting curve analysis and an example thereof according to an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be included. Preferably, the digital melting curve analysis and visualization system (10) based on ultra-precision temperature resolution may be a system capable of detecting microsatellite instability (MSI) with high precision.

[0238] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can analyze the microsatellite structure of representative MSI markers such as BAT26 and CAT25.

[0239] Preferably, BAT26 is a marker containing 26 A repeat sequences and can be used to determine the presence of Mismatch Repair (MMR) defects in regions where replication errors frequently occur.

[0240] Preferably, CAT25 is a marker containing 25 T repeat sequences, which likewise has a high replication error frequency and can be an important indicator for determining MSI status.

[0241] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can construct an MSI standard sample library by combining normal (MSS) sequences and MSI templates in which replication errors are artificially inserted. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can generate and analyze an MSI Tm histogram by performing digital PCR-based melting curve analysis. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide the MSI status for an unknown sample in the form of a quantitative histogram.

[0242] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can distinguish precise MSI based on minute Tm changes through the high-resolution characteristics of digital melting analysis, unlike the conventional capillary electrophoresis-based size analysis method.

[0243] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can visualize the BAT26 and CAT25 analysis results of MSI-H (Microsatellite Instability-High) patients, and can quantitatively confirm the presence of instability by clearly observing the movement and shape change of the melting curve, especially in CAT25. Preferably, the analysis results of the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can be clinically utilized in predicting the response to immune checkpoint inhibitors such as Pembrolizumab, diagnosing hereditary colorectal cancer (HNPCC), and selecting candidates for immunotherapy.

[0244] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can be applied as a diagnostic platform superior in terms of sensitivity and precision compared to existing methods.

[0245] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can monitor the dissociation process of the DNA double helix structure in real time according to temperature changes and detect minute differences in the DNA sequence.

[0246] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) provides an ultra-precision temperature resolution of 0.1°C or less, enabling high-sensitivity analysis capable of detecting even single base mutations. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) can provide much more precise results than conventional bulk analysis methods by enabling analysis at the individual DNA molecule level in combination with digital PCR technology.

[0247] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) includes an algorithm for quantitative evaluation of the MSI state, and can comprehensively analyze various parameters such as the shape of the melting curve, the location of the peak, and the slope of the curve.

[0248] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization system (10) provides a visualization interface that allows the MSI detection results to be intuitively understood, so that a clinician can easily interpret the results and use them for diagnosis.

[0249] According to one embodiment, a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include at least one of a data input step, a temperature resolution correction step, and a high-resolution Tm data processing step. The data input step may receive raw melting data collected from a real-time digital PCR platform. The temperature resolution correction step may convert the measured temperature data into high-resolution temperature data. The high-resolution Tm data processing step may precisely extract the interpolated Tm data.

[0250] According to another embodiment, the data input step may include a real-time digital PCR platform data interface step that collects data in conjunction with a real-time digital PCR platform. This allows for the efficient collection of data for digital melting curve analysis and visualization based on ultra-precision temperature resolution.

[0251] According to another embodiment, the temperature resolution correction step may include a linear interpolation application step that generates additional temperature points using a linear interpolation method based on the measured temperature intervals and converts them into a high-resolution curve of 0.1°C or less. Through this linear interpolation application, ultra-precision temperature resolution can be achieved to improve the analysis accuracy of the digital melting curve.

[0252] According to another embodiment, the temperature resolution correction step may include a derivative curve generation step that improves the precision of Tm extraction by smoothly generating the first derivative (-dF / dT) curve of each well. Through such derivative curve generation, the analysis accuracy of the digital melting curve can be increased.

[0253] According to another embodiment, the high-resolution Tm data processing step may include a Tm data segmentation step that generates a Tm data histogram by segmenting the x-axis into temperature units smaller than the temperature units of the raw input data. Through this segmentation process, the precision can be increased by analyzing the data in finer temperature units.

[0254] According to another embodiment, the high-resolution Tm data processing step may include a well count calculation step that represents the number of wells with the corresponding Tm on the y-axis of a histogram. Through this well count calculation, the distribution of wells where melting occurred at a specific temperature can be accurately identified.

[0255] According to another embodiment, the high-resolution Tm data processing step may include a high-resolution visualization step that has a resolution 3 to 5 times higher than the existing resolution and visualizes Tm distributions that are close to each other so as to be clearly distinguishable. Through this high-resolution visualization, close Tm distributions that were difficult to distinguish with existing methods can be analyzed by distinguishing them more clearly.

[0256] According to another embodiment, the high-resolution Tm data processing step may include a PCR control peak reference alignment step, which sets the PCR control peak used in the melting analysis as a reference value and reference-calibrates the Tm values ​​of the entire sample based on this reference value to reduce errors between experiments or equipment and ensure quantitative accuracy for comparison between data. Through this PCR control peak reference alignment step, variations that may occur in various experimental environments can be minimized, and more accurate analysis results can be obtained.

[0257] According to another embodiment, the method may further include an analysis software execution step for executing a high-resolution correction algorithm and a visualization UI in the analysis software. Such analysis software execution can improve the efficiency of the ultra-precision temperature resolution-based digital melting curve analysis and visualization method.

[0258] According to another embodiment, the analysis software execution step may include a data smoothing step that processes data smoothly to improve analysis accuracy. This data smoothing step can contribute to increasing the accuracy of the ultra-precision temperature resolution-based digital melting curve analysis and visualization method.

[0259] According to another embodiment, the analysis software execution step may include a noise removal step that improves data quality by removing noise generated in the data. Through this noise removal step, the accuracy of the digital melting curve analysis based on ultra-precision temperature resolution can be increased.

[0260] According to another embodiment, the analysis software execution step may include a threshold filtering step that filters data based on a set threshold. Through this threshold filtering, unnecessary data can be removed and only meaningful data can be selectively processed during the process of digital melting curve analysis and visualization based on ultra-precision temperature resolution.

[0262] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include a data input step (1000), a temperature resolution correction step (2000), and a high-resolution Tm data processing step (3000).

[0263] Preferably, the data input step (1000) may be a step of receiving raw melting data collected from a real-time digital PCR platform. Preferably, in the data input step (1000), data regarding fluorescence signal intensity and temperature changes generated from a digital PCR instrument may be collected in a time-series form. Preferably, this raw melting data may include changes in fluorescence signals occurring during the process of DNA double strands separating into single strands. Preferably, the data input step (1000) may support data files of various formats to process data generated from different PCR instruments. Preferably, the data input step (1000) may include a function to verify the integrity of the input data and identify outliers. Preferably, the temperature resolution correction step (2000) may be a step of converting the measured temperature data into high-resolution temperature data. Preferably, in the temperature resolution correction step (2000), temperature data at intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C provided by a general PCR instrument can be corrected to an ultra-precision temperature resolution of 0.1°C or less. Preferably, this correction process can be performed using mathematical algorithms such as linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression. Preferably, the temperature resolution correction step (2000) may include a correction algorithm for correcting the temperature sensor error of the instrument.

[0265] Preferably, the data input step (1000), the temperature resolution correction step (2000), and the high-resolution Tm data processing step (3000) are executed sequentially to provide digital melting curve analysis results based on ultra-precision temperature resolution. Preferably, these analysis results can be visualized through a graphical user interface and provided in a form that the user can intuitively understand. Preferably, the visualized results can be expressed in various forms, such as the original melting curve, the normalized melting curve, and the differential melting curve. Preferably, the present method can provide high accuracy and reproducibility in various PCR applications, particularly in SNP (Single Nucleotide Polymorphism) analysis, methylation analysis, and mutation detection. Preferably, through ultra-precision temperature resolution, DNA sequences with similar Tm values ​​that were difficult to distinguish with conventional methods can be clearly distinguished. Preferably, the present method can be linked with a cloud-based system to enable the processing of large-scale data and the sharing of analysis results.

[0266] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include a data input step (1000) and a real-time digital PCR platform data interface step (1100). Preferably, the data input step (1000) may be a step of receiving raw melting data collected from a real-time digital PCR platform. Preferably, the data input step (1000) may collect data measured according to temperature changes regarding changes in fluorescence signals occurring during the process of separating DNA double strands into single strands after a digital PCR reaction. Preferably, the data input step (1000) may be designed to process various forms of raw data formats. Preferably, the data input step (1000) may include a real-time digital PCR platform data interface step (1100).

[0267] Preferably, the real-time digital PCR platform data interface step (1100) can collect data by linking with the real-time digital PCR platform. Preferably, the real-time digital PCR platform data interface step (1100) can collect data in real time by directly connecting to digital PCR equipment from various manufacturers through an API (Application Programming Interface). Preferably, the real-time digital PCR platform data interface step (1100) can provide a flexible data collection environment by supporting various communication protocols such as USB, Ethernet, Bluetooth, and Wi-Fi. Preferably, the real-time digital PCR platform data interface step (1100) can ensure reliable data collection by including an algorithm that verifies the integrity of the collected data. Preferably, the data input step (1000) may include the real-time digital PCR platform data interface step (1100).

[0268] Preferably, the real-time digital PCR platform data interface step (1100) can collect data with an ultra-precision temperature resolution of 0.1°C or less, so that changes in the fluorescence signal due to minute temperature changes can be accurately captured. Preferably, the real-time digital PCR platform data interface step (1100) can convert the collected data into a normalized format to ensure compatibility in subsequent analysis processes. Preferably, the real-time digital PCR platform data interface step (1100) may include a function to filter noise generated during the data collection process in real time.

[0269] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include an interpolation application step (2100). Preferably, the interpolation application step (2100) may generate additional temperature points using a linear interpolation method based on the measured temperature intervals and convert them into a high-resolution curve with a unit of 0.1°C or less. Preferably, the interpolation application step (2100) may mathematically generate additional data points with a 0.1°C interval between temperature data points with intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C that are basically provided by the experimental equipment. Preferably, the interpolation application step (2100) may calculate a fluorescence signal value corresponding to a temperature value located between two adjacent measured temperature points by assuming a linear relationship between them. Preferably, the interpolation application step (2100) can improve the temperature resolution by more than three times to more accurately capture minute changes in the melting curve. Preferably, the temperature resolution correction step (2000) may include the interpolation application step (2100). Preferably, the temperature resolution correction step (2000) can convert the measured temperature data into high-resolution temperature data. Preferably, the temperature resolution correction step (2000) can receive and process raw fluorescence data and temperature data measured by a real-time PCR instrument. Preferably, the temperature resolution correction step (2000) can selectively apply various interpolation methods, such as spline interpolation, polynomial interpolation, or Gaussian process regression-based interpolation, in addition to linear interpolation.Preferably, the high-resolution temperature data generated in the interpolation application step (2100) can be utilized in subsequent analysis steps to accurately identify the location of melting peaks, analyze the pattern of fluorescence signal change according to minute temperature changes, and improve the resolution of complex melting curves with multiple peaks. Preferably, the interpolation application step (2100) can play an important role in detecting genetic variations distinguished by small temperature differences, particularly in SNP (Single Nucleotide Polymorphism) analysis or methylation analysis.

[0270] Preferably, in the interpolation application step (2100), linear interpolation can be implemented using the following mathematical formula: y = y₁ + ((x - x₁) / (x₂ - x₁)) * (y₂ - y₁), where x is the temperature value to be interpolated, y is the calculated fluorescence signal value, and (x₁, y₁) and (x₂, y₂) may each be two adjacent measurement data points. Preferably, the interpolation application step (2100) can be automated through a computer program to efficiently process a large amount of data.

[0271] An embodiment of a digital melting curve analysis and visualization method based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a derivative curve generation step (2200). Preferably, the derivative curve generation step (2200) can improve the precision of Tm extraction by smoothly generating a first derivative (-dF / dT) curve of each well. Preferably, the derivative curve generation step (2200) can generate a first derivative curve by calculating the rate of change of the fluorescence signal according to the temperature change. Preferably, the derivative curve generation step (2200) can generate a smooth curve with noise removed by applying mathematical techniques such as linear interpolation, spline interpolation, polynomial interpolation, or interpolation based on Gaussian process regression. Preferably, the smooth first derivative (-dF / dT) curve generated in the derivative curve generation step (2200) can play an important role in identifying the exact location of Tm.

[0272] Preferably, the temperature resolution correction step (2000) may include a derivative curve generation step (2200). Preferably, the temperature resolution correction step (2000) may convert the measured temperature data into high-resolution temperature data. Preferably, the temperature resolution correction step (2000) may provide more precise temperature data by resolving discontinuities or low-resolution issues in the initially measured temperature data. Preferably, in the derivative curve generation step (2200), noise may be minimized by optimizing the spacing between adjacent temperature points when calculating the rate of change of the fluorescence signal for each well. Preferably, the derivative curve generation step (2200) may mitigate abrupt fluctuations in the data by applying a moving average filter. Preferably, the smooth curve generated in the derivative curve generation step (2200) may improve the extraction precision of Tm by enabling more accurate identification of peak locations.

[0273] Preferably, the derivative curve generation step (2200) may selectively apply an algorithm optimized according to the characteristics of various DNA samples. Preferably, the derivative curve generation step (2200) may derive optimal results by applying different smoothing parameters depending on the GC content or length of the sample. Preferably, the derivative curve generation step (2200) may adapt to various experimental conditions by adjusting parameters in real time. Preferably, the smooth first derivative (-dF / dT) curve generated in the derivative curve generation step (2200) may be particularly useful in advanced molecular diagnostic techniques such as multiplex PCR or HRM (High Resolution Melting) analysis. Preferably, the derivative curve generation step (2200) may improve the accuracy of SNP (single nucleotide polymorphism) analysis by enabling clear distinction of each peak even in complex melting profiles where multiple peaks exist in close proximity.

[0274] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include a Tm data segmentation step (3100). Preferably, the Tm data segmentation step (3100) may generate a Tm data histogram by segmenting the x-axis into temperature units smaller than the temperature units of the raw input data. Preferably, in the Tm data segmentation step (3100), if the temperature units of the raw input data are intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C, the x-axis may be segmented into units of 0.1°C or smaller than these. Preferably, through such fine temperature unit segmentation, peaks with fine temperature differences that were difficult to distinguish using conventional methods can be effectively separated and visualized.

[0275] Preferably, the histogram generated in the Tm data segmentation step (3100) can display the frequency of the fluorescence signal corresponding to each temperature segment on the y-axis. Preferably, through this histogram, the dissociation phenomenon of the DNA double strand occurring at a specific temperature can be observed more precisely.

[0276] Preferably, the high-resolution Tm data processing step (3000) may include a Tm data segmentation step (3100). Preferably, the high-resolution Tm data processing step (3000) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing step (3000) can remove noise or background signals that may occur in the raw data and selectively extract only the actual meaningful Tm data. Preferably, the histogram generated in the Tm data segmentation step (3100) can be represented through various visualization techniques. Preferably, it can be converted into a form such as a heat map, 3D surface map, or contour map to more clearly identify patterns or anomalies in the data. Preferably, the Tm data segmentation step (3100) may include a process of finding the optimal resolution by applying various segment sizes. Preferably, the optimal segment size suitable for the characteristics of the data can be determined through an automated algorithm, thereby improving the accuracy of the analysis.

[0277] Preferably, in the Tm data segmentation step (3100), statistical analysis can be performed on the segmented data to evaluate the statistical significance of each peak. Preferably, this allows distinguishing between peaks with actual biological significance and peaks caused by noise.

[0278] An embodiment of a digital melting curve analysis and visualization method based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a well count calculation step (3200).

[0279] Preferably, the well count calculation step (3200) can represent the number of wells with the corresponding Tm on the y-axis of a histogram. Preferably, the well count calculation step (3200) can accurately calculate the number of wells with the corresponding Tm for each temperature range and visualize it in the form of a histogram. Preferably, the well count calculation step (3200) can aggregate the number of wells with the corresponding Tm value for each temperature range divided by ultra-precision temperature resolution of 0.1°C or less, such as 0.1°C, 0.05°C, or 0.01°C. Preferably, the well count calculation step (3200) can be applied to microplates of various specifications, such as 8 strips, 96-well plates, 384-well plates, 1536-well plates, or plates with more than 20,000 wells.

[0280] Preferably, the high-resolution Tm data processing step (3000) may include a well count calculation step (3200). Preferably, the high-resolution Tm data processing step (3000) may precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing step (3000) may determine the Tm value by accurately identifying the point where the rate of change of the fluorescence signal measured at each well is maximum. Preferably, the high-resolution Tm data processing step (3000) may improve the signal-to-noise ratio by applying a noise filtering algorithm.

[0281] Preferably, the histogram generated in the well count calculation step (3200) allows for an intuitive understanding of the distribution pattern for specific Tm values, which can be utilized for the analysis of genetic variation or mutations between samples. Preferably, the well count calculation step (3200) can automatically identify major peaks in the histogram and evaluate the statistical significance of each peak through a multiple peak detection algorithm. Preferably, the well count calculation step (3200) can calculate the sum of well counts within a user-specified temperature range to analyze the proportion of samples belonging to a specific Tm range. Preferably, the well count calculation step (3200) can visualize the spatial distribution pattern on the microplate by linking the location information of the corresponding well with the Tm value of each well. Preferably, the well count calculation step (3200) can track changes in the Tm distribution over time and be utilized for the analysis of reaction kinetics. Preferably, the well count calculation step (3200) can distinguish different Tm ranges using various color codes, thereby facilitating the interpretation of complex multiplex PCR results.

[0282] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include a PCR control peak-based alignment step (3300).

[0283] Preferably, the PCR control peak reference alignment step (3300) sets the PCR control peak used in the melting analysis as a reference value and reference calibrates the Tm values ​​of the entire sample based on this to reduce errors between experiments or equipment and ensure quantitative accuracy for comparison between data. Preferably, the PCR control peak reference alignment step (3300) can adjust the melting curves of all other samples by utilizing the melting peak of the standard control sample included in each experiment set as a reference point. Preferably, the high-resolution Tm data processing step (3000) may include the PCR control peak reference alignment step (3300).

[0284] Preferably, in the PCR control peak reference alignment step (3300), a synthetic oligonucleotide or standard DNA sample having a known Tm value may be used as the PCR control peak. Preferably, while there may be slight variations in this PCR control peak depending on experimental conditions or equipment status, its relative position may be maintained constant. Preferably, the PCR control peak reference alignment step (3300) may derive a correction factor by calculating the difference between the theoretical Tm value of this PCR control peak and the actual measured Tm value. Preferably, the correction factor derived in the PCR control peak reference alignment step (3300) may be applied uniformly to all samples within the same experimental set to adjust the Tm value of the entire dataset. Preferably, this alignment process may enable direct comparison between data obtained from different experimental dates, different PCR instruments, or different laboratories. Preferably, the PCR control peak reference alignment step (3300) can be applied particularly effectively to melting curve data with an ultra-precision temperature resolution of 0.1°C or less. Preferably, the PCR control peak reference alignment step (3300) can also perform non-linear correction according to the temperature range by using multiple PCR control peaks simultaneously. Preferably, this multiple reference point alignment method can effectively correct non-linear temperature deviations that may occur over a wide temperature range.

[0285] Preferably, the PCR control peak reference alignment step (3300) can automatically recognize the PCR control peak and perform the alignment process through an automated algorithm. Preferably, this algorithm can detect a peak within a specific temperature range in the first derivative curve of the melting curve and determine the exact location of the peak using methods such as Gaussian fitting. Preferably, the difference between the location of the PCR control peak determined in this way and the reference location can be calculated and applied to all data points. Preferably, the melting curve data aligned through the PCR control peak reference alignment step (3300) can provide high reliability and reproducibility in subsequent analysis steps. Preferably, this alignment process can play an important role in applications such as SNP analysis, methylation analysis, or gene variant detection, where minute Tm differences between multiple samples must be detected. Preferably, the PCR control peak reference alignment step (3300) can ensure data consistency, which is essential in large-scale clinical studies or diagnostic applications, by minimizing inter-experiment variability.

[0286] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include an analysis software execution step (4000). Preferably, the analysis software execution step (4000) may include a process of executing a high-resolution correction algorithm and a visualization UI in the analysis software. Preferably, the analysis software may be a software program that can be run on various electronic devices, including a computer. Preferably, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include an analysis software execution step (4000). Preferably, the high-resolution correction algorithm may include a mathematical model and a calculation method to improve the precision of the collected temperature data. Preferably, this algorithm may be designed to remove noise that may occur during the temperature measurement process and to detect fine patterns of temperature change. Preferably, the high-resolution correction algorithm may improve the quality of the temperature data by utilizing signal processing techniques such as Fourier transform, wavelet analysis, and Kalman filtering.

[0287] Preferably, the visualization UI (user interface) can display the analyzed melting curve data in a graphical form that the user can intuitively understand. Preferably, the visualization UI can provide various visualization methods, such as 2D graphs, 3D modeling, heat maps, and contour plots. Preferably, through the visualization UI, the user can zoom in on a specific section of the melting curve, compare the melting curves of multiple samples, or intensively observe changes within a specific temperature range.

[0288] Preferably, the analysis software execution step (4000) can provide a real-time data processing function. Preferably, through this function, the user can monitor and analyze changes in the melting curve in real time while the experiment is in progress. Preferably, the real-time processing function can receive data directly from the data collection device, process it without delay, and visualize it.

[0289] Preferably, the analysis software execution step (4000) may include data export and report generation functions. Preferably, the user may export the analyzed data in various formats such as CSV, Excel, PDF, etc., or generate an automated report. Preferably, the generated report may include a melting curve graph, statistical analysis results, sample information, etc.

[0290] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include a data smoothing step (4100).

[0291] Preferably, the data smoothing step (4100) can improve analysis accuracy by smoothing the data. Preferably, the data smoothing step (4100) can remove noise or irregular fluctuations that may occur in the melting curve data to enable the identification of clearer patterns. Preferably, the data smoothing step (4100) can be implemented using various mathematical algorithms such as the moving average method, Gaussian filter, and Savitzky-Golay filter. Preferably, the data smoothing step (4100) can operate in a manner that removes only unnecessary fluctuations while preserving important characteristics of the original data. Preferably, the analysis software execution step (4000) may include the data smoothing step (4100). Preferably, the analysis software execution step (4000) can execute a high-resolution correction algorithm and a visualization UI in the analysis software. Preferably, the analysis software execution step (4000) can provide an interface that allows the user to adjust the intensity of the data smoothing.

[0292] Preferably, the data smoothing step (4100) enables more precise analysis, particularly in sections with minute temperature changes, so that even small changes can be detected without being missed. Preferably, the data smoothing step (4100) can improve the accuracy of various analysis tasks, such as melting point identification, transition temperature detection, and double peak analysis.

[0293] Preferably, the data smoothing step (4100) can automatically suggest smoothing parameters optimized for various sample types and experimental conditions. Preferably, the data smoothing step (4100) can calculate the statistical reliability of the processed data and provide it to the user.

[0294] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include a noise removal step (4200). Preferably, the noise removal step (4200) can improve data quality by removing noise occurring in the data. Preferably, the noise removal step (4200) can identify and remove noise caused by random signal fluctuations, electrical interference, or measurement errors occurring in the digital melting curve data. Preferably, the noise removal step (4200) can effectively remove noise by utilizing various signal processing algorithms such as a moving average filter, a Gaussian filter, or a wavelet transform. Preferably, the noise removal step (4200) can selectively remove only unnecessary signal fluctuations while preserving important characteristics of the original data.

[0295] Preferably, the noise removal step (4200) can automatically adjust the filtering intensity according to the characteristics of the data by applying a filtering technique. Preferably, the noise removal step (4200) can identify and correct abnormal data points using a statistical outlier detection method. Preferably, the noise removal step (4200) can remove noise caused by electrical interference or temperature fluctuations occurring during the measurement process while preserving minute changes in the melting curve. Preferably, the noise removal step (4200) can improve the signal-to-noise ratio (SNR) to increase the accuracy of subsequent analysis. Preferably, the noise removal step (4200) can process data in real time to provide immediate analysis results.

[0296] Preferably, the analysis software execution step (4000) may include a noise removal step (4200). Preferably, the analysis software execution step (4000) may execute a high-resolution correction algorithm and a visualization UI to enable accurate analysis and intuitive visualization of melting curve data. Preferably, the analysis software execution step (4000) may derive more accurate analysis results based on the data refined through the noise removal step (4200). Preferably, the noise removal step (4200) may provide data quality evaluation metrics so that the user can quantitatively verify the noise removal effect. Preferably, the noise removal step (4200) may provide a function to visually compare the original data and the noise-removed data. Preferably, the noise removal step (4200) may provide parameter setting options that allow the user to adjust the noise removal intensity.

[0297] An embodiment of a method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution is described in detail. According to an embodiment of the present invention, the method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution may include a threshold filtering step (4300).

[0298] Preferably, the threshold filtering step (4300) can filter data based on a set threshold. Preferably, the threshold filtering step (4300) can remove noise or unnecessary signals from the melting curve data based on a specific threshold defined by the user or a threshold automatically calculated by the system. Preferably, the threshold filtering step (4300) can be applied to various parameters such as the intensity of the fluorescence signal, the rate of temperature change, or the signal-to-noise ratio. Preferably, the threshold filtering step (4300) can contribute to improving the quality of the data and increasing the accuracy of the analysis. Preferably, the analysis software execution step (4000) may include the threshold filtering step (4300). Preferably, the analysis software execution step (4000) can execute high-resolution correction algorithms and visualization UIs in the analysis software. Preferably, the analysis software execution step (4000) can manage the entire process of processing and analyzing the melting curve data. Preferably, the threshold filtering step (4300) can dynamically adjust the threshold by utilizing various algorithms. Preferably, the threshold filtering step (4300) can automatically determine the optimal threshold by analyzing the statistical distribution of the data. Preferably, the threshold filtering step (4300) can provide a function to manually adjust the threshold through a user interface.

[0299] Preferably, the threshold filtering step (4300) may include a function to visually verify the effect of filtering by comparing the filtered data with the original data. Preferably, the threshold filtering step (4300) may update the filtering results in real time to support the user in finding the optimal threshold.

[0300] Preferably, the threshold filtering step (4300) can perform multiple stages of filtering by setting an upper Tm threshold or a lower Tm threshold. Preferably, the threshold filtering step (4300) can enable more precise data filtering by applying different thresholds for each temperature range. Preferably, the threshold filtering step (4300) can provide a function to automatically save filtered data and compare it with previous analysis results. Preferably, the threshold filtering step (4300) can record the data removed during the filtering process separately so that it can be reviewed when necessary.

[0301] Referring to FIG. 22, FIG. 22 is a conceptual flowchart of a digital melting curve analysis and visualization method based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. According to an embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a data input step (1000), a temperature resolution correction step (2000), and a high-resolution Tm data processing step (3000).

[0302] Preferably, the data input step (1000) may be a step of receiving raw melting data collected from a real-time digital PCR platform. Preferably, the data input step (1000) may collect data based on fluorescence signal intensity and temperature changes generated from a digital PCR device.

[0303] Preferably, the raw melting data collected in the data input step (1000) may include changes in the fluorescent signal that occur during the process of separating the double strand of a DNA or RNA sample into a single strand.

[0304] Preferably, the data input step (1000) can process fluorescence signal data measured simultaneously in multiple wells in parallel.

[0305] Preferably, the temperature resolution correction step (2000) may be a step of converting measured temperature data into high-resolution temperature data. Preferably, the temperature resolution correction step (2000) may correct existing temperature data in units of 0.2°C, 0.3°C, 0.5°C, or 1.0°C to an ultra-precision temperature resolution of 0.1°C or less, e.g., 0.1°C, 0.05°C, or 0.01°C.

[0306] Preferably, the temperature resolution correction step (2000) can improve the resolution of the temperature data by utilizing linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression-based interpolation.

[0307] Preferably, the temperature resolution correction step (2000) can correct errors that may occur in the temperature sensor of the PCR equipment to provide accurate temperature data. Preferably, the high-resolution Tm data processing step (3000) may be a step of precisely extracting interpolated Tm data.

[0308] Preferably, the high-resolution Tm data processing step (3000) can calculate the first derivative of the fluorescence signal to accurately identify the inflection point of the melting curve.

[0309] Preferably, the high-resolution Tm data processing step (3000) can individually analyze each peak when multiple Tm peaks are present to confirm the presence of a complex DNA structure or multiple targets.

[0310] Preferably, the high-resolution Tm data processing step (3000) can improve the signal-to-noise ratio (SNR) by applying a noise filtering algorithm.

[0311] Preferably, according to one embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may further include a data visualization step. Preferably, the data visualization step may represent high-resolution processed melting curve data as a 2D or 3D graph so that a user can intuitively understand the analysis results. Preferably, according to one embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may further include a data storage and sharing step.

[0312] Preferably, the data storage and sharing step can store the analyzed high-resolution Tm data in a normalized format and enable it to be shared with other researchers through a cloud-based platform.

[0313] Preferably, according to one embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may further include a statistical analysis step.

[0314] Preferably, the statistical analysis step can evaluate the reproducibility and reliability of the experiment by calculating the distribution, mean, standard deviation, etc. of Tm data obtained from multiple samples. Preferably, according to one embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may further include an automated quality control step.

[0315] Preferably, the automated quality control step can automatically evaluate the quality of the analyzed data, identify outliers, and notify the user.

[0316] Referring to FIG. 23, FIG. 23 is a conceptual flowchart of a data input step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a data input step (1000) and a real-time digital PCR platform data interface step (1100).

[0317] Preferably, according to one embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a data input step (1000). Preferably, the data input step (1000) may be a step of receiving raw melting data collected from a real-time digital PCR platform.

[0318] Preferably, the data input step (1000) can provide a flexible interface capable of receiving melting data generated from various types of digital PCR platforms. Preferably, the data input step (1000) can process raw data representing changes in fluorescence signal intensity according to temperature changes.

[0319] Preferably, the data input step (1000) can support data in various file formats (CSV, TXT, XML, etc.) to improve user convenience.

[0320] Preferably, the data input step (1000) can optimize the input speed by utilizing a parallel processing algorithm for large-scale data processing.

[0321] Preferably, according to one embodiment of the present invention, the data input step (1000) may include a real-time digital PCR platform data interface step (1100). Preferably, the real-time digital PCR platform data interface step (1100) may collect data in conjunction with a real-time digital PCR platform.

[0322] Preferably, the real-time digital PCR platform data interface step (1100) can communicate directly with digital PCR equipment from various manufacturers via an Application Programming Interface (API). Preferably, the real-time digital PCR platform data interface step (1100) can ensure the accuracy of temperature measurement during the data collection process with a resolution of 0.1°C or less. Preferably, the real-time digital PCR platform data interface step (1100) can monitor data in real time and immediately detect outliers to provide notifications to the user. Preferably, the real-time digital PCR platform data interface step (1100) can maintain the security of sensitive genetic information by using an encrypted data transmission protocol. Preferably, the interaction between the real-time digital PCR platform data interface step (1100) and the data input step (1000) may include a checksum verification process to ensure data integrity. Preferably, the data input step (1000) can perform an initial quality assessment of the input raw melting data to determine whether it is suitable for subsequent analysis. Preferably, the data input step (1000) can have the ability to process fluorescence data collected simultaneously from multiple channels.

[0323] Preferably, the data input step (1000) can provide a function to selectively extract only data within a specific temperature range defined by the user.

[0324] Preferably, the data input step (1000) can automatically extract and store metadata (experimental conditions, time, equipment information, etc.) of the input data.

[0325] Referring to FIG. 24, FIG. 24 is a conceptual flowchart of a temperature resolution correction step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a temperature resolution correction step (2000) and an interpolation application step (2100).

[0326] Preferably, the temperature resolution correction step (2000) can convert the measured temperature data into high-resolution temperature data. Preferably, the temperature resolution correction step (2000) can improve the resolution of the temperature data obtained from existing measuring equipment to enable more precise analysis. Preferably, the temperature resolution correction step (2000) can reduce discontinuities in the measured temperature data and generate a smoother temperature curve. Preferably, the temperature resolution correction step (2000) can overcome the resolution limitations of the temperature data caused by the physical limitations of the measuring equipment using a software method. Preferably, according to one embodiment of the present invention, the temperature resolution correction step (2000) may include an interpolation application step (2100). Preferably, the interpolation application step (2100) can generate additional temperature points using a linear interpolation method based on the measured temperature interval to convert the temperature into a high-resolution curve of 0.1°C or less. Preferably, the interpolation application step (2100) can mathematically calculate and insert new temperature points between two consecutive measured temperature points. Preferably, the interpolation application step (2100) can proportionally calculate the values ​​between them by assuming a linear relationship between two adjacent temperature points. For example, the interpolation application step (2100) can provide a temperature resolution that is at least 2 to 100 times denser by generating new data points at 0.1°C intervals between temperature data points measured at intervals of 0.2°C, 0.3°C, 0.5°C, or 1.0°C. In one embodiment, the interpolation application step (2100) can perform interpolation at finer temperature intervals, such as 0.05°C or 0.01°C, as needed. Preferably, the interpolation application step (2100) can generate a smoother and more continuous melting curve based on the interpolated temperature data.Preferably, the high-resolution temperature data generated in the interpolation application step (2100) can more accurately capture changes in the fluorescence signal due to minute temperature changes in the subsequent analysis step. Preferably, the high-resolution temperature data generated through the interpolation application step (2100) can help to more clearly identify minute inflection points or peaks of the melting curve. Preferably, the interpolation application step (2100) is performed in real time so that high-resolution data can be provided immediately upon measurement.

[0327] Preferably, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution can perform differential analysis of the melting curve based on high-resolution temperature data generated through the temperature resolution correction step (2000). Preferably, the high-resolution temperature data can be utilized to more accurately analyze the melting characteristics of nucleic acids, such as DNA or RNA. Preferably, the high-resolution data generated through the temperature resolution correction step (2000) can be intuitively presented to the user through various visualization techniques.

[0328] Referring to FIG. 25, FIG. 25 is a conceptual flowchart of a temperature resolution correction step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a temperature resolution correction step (2000) and a derivative curve generation step (2200).

[0329] Preferably, the temperature resolution correction step (2000) can convert the measured temperature data into high-resolution temperature data. Preferably, the temperature resolution correction step (2000) can receive and process raw temperature data measured in real time. Preferably, the temperature resolution correction step (2000) can generate continuous high-resolution temperature data by compensating for discontinuities in temperature data caused by physical limitations of the measuring equipment. Preferably, the temperature resolution correction step (2000) can improve the resolution of the temperature data by utilizing linear interpolation, spline interpolation, polynomial interpolation, or interpolation based on Gaussian process regression.

[0330] Preferably, according to one embodiment of the present invention, the temperature resolution correction step (2000) may include a derivative curve generation step (2200). Preferably, the derivative curve generation step (2200) can improve the precision of Tm extraction by smoothly generating a first derivative (-dF / dT) curve for each well. Preferably, the derivative curve generation step (2200) can calculate a first derivative curve based on high-resolution converted temperature data and fluorescence signal data. Preferably, the derivative curve generation step (2200) can apply signal processing techniques such as moving average filtering or Gaussian smoothing to minimize noise. Preferably, the derivative curve generation step (2200) can generate independent derivative curves for each well to facilitate comparative analysis between samples. Preferably, the derivative curve generation step (2200) can accurately extract Tm values ​​by automatically detecting peak values ​​from the generated derivative curves. Preferably, the derivative curve generation step (2200) can distinguish between the main peak and the volumetric peak by analyzing the height and width of each peak when multiple peaks exist. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization method may additionally include a data visualization step in addition to the temperature resolution correction step (2000) and the derivative curve generation step (2200). Preferably, the data visualization step can represent the generated high-resolution melting curve and derivative curve in the form of a graph that can be intuitively understood. Preferably, the data visualization step can provide a function that allows simultaneous comparison of curves of multiple samples using various colors and markers.

[0331] Preferably, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may additionally include a database management step for storing and managing analysis results. Preferably, the database management step may store the analyzed melting curve data in a structured form to facilitate future searching and comparative analysis.

[0332] Referring to FIG. 26, FIG. 26 is a conceptual flowchart of a high-resolution Tm data processing step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a high-resolution Tm data processing step (3000) and a Tm data segmentation step (3100).

[0333] Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization method may be a method for analyzing and visualizing the melting characteristics of nucleic acid molecules, such as DNA or RNA, of a biological sample at high resolution. Preferably, the high-resolution Tm data processing step (3000) may be a process for precisely extracting interpolated Tm data. Preferably, the high-resolution Tm data processing step (3000) may receive and process raw data obtained from a real-time PCR instrument or other molecular analysis instrument.

[0334] Preferably, the high-resolution Tm data processing step (3000) can analyze the melting characteristics of nucleic acid molecules by measuring changes in fluorescence signals according to temperature. Preferably, the high-resolution Tm data processing step (3000) may include a preprocessing step for removing background signals and filtering noise. Preferably, the high-resolution Tm data processing step (3000) may include a Tm data segmentation step (3100). Preferably, the Tm data segmentation step (3100) may be a process of generating a Tm data histogram by segmenting the x-axis into temperature units smaller than the temperature units of the raw input data. Preferably, the temperature units used in the Tm data segmentation step (3100) may have an ultra-precision temperature resolution of 0.1°C or less.

[0335] Preferably, the Tm data interval step (3100) can detect minute Tm differences by setting a temperature interval that is much more precise than the existing temperature resolution of 0.2℃, 0.3℃, 0.5℃, or 1.0℃.

[0336] Preferably, the Tm data interval step (3100) can calculate the frequency of data points corresponding to each temperature interval and express them in the form of a histogram.

[0337] Preferably, the histogram generated in the Tm data interval step (3100) can visually represent the melting frequency of nucleic acid molecules at a specific temperature.

[0338] Preferably, the Tm data segmentation step (3100) can provide a resolution that allows each peak to be clearly distinguished even in complex melting curves with multiple peaks. Preferably, the Tm data segmentation step (3100) can secure the precision required to detect single nucleotide polymorphisms (SNPs) or minute genetic variations. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization method can be utilized in various fields such as clinical diagnosis, forensic analysis, and microbial identification.

[0339] Referring to FIG. 27, FIG. 27 is a conceptual flowchart of a high-resolution Tm data processing step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a high-resolution Tm data processing step (3000) and a well count calculation step (3200).

[0340] Preferably, the high-resolution Tm data processing step (3000) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing step (3000) may include a process of extracting Tm from raw fluorescence data obtained after a digital PCR reaction. Preferably, the high-resolution Tm data processing step (3000) can accurately identify the inflection point of the melting curve by calculating the derivative value of the fluorescence signal. Preferably, the high-resolution Tm data processing step (3000) can improve the signal-to-noise ratio by applying a noise filtering algorithm.

[0341] Preferably, the high-resolution Tm data processing step (3000) can correct temperature deviations that may occur between devices through a temperature correction algorithm. Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing step (3000) may include a well count calculation step (3200). Preferably, the well count calculation step (3200) can represent the number of wells with the corresponding Tm on the y-axis of a histogram.

[0342] Preferably, the well count calculation step (3200) can visualize the distribution by calculating the number of wells with the same Tm for each temperature range.

[0343] Preferably, the well count calculation step (3200) allows the user to adjust the temperature range from 0.01℃ to 1℃, thereby allowing the resolution of the histogram to be adjusted according to the purpose of analysis. Preferably, the well count calculation step (3200) can generate a normalized histogram based on the calculated well count data to facilitate comparison between different experiments. Preferably, the well count calculation step (3200) can automatically identify major Tm peaks in the histogram by applying a multi-peak detection algorithm. Preferably, the well count calculation step (3200) can evaluate the reliability of the analysis results by calculating the statistical characteristics (mean, standard deviation, kurtosis, skewness, etc.) of the identified peaks. Preferably, the well count calculation step (3200) can facilitate further analysis by providing a function to export the calculated well count data in various formats such as CSV and Excel.

[0344] Preferably, the high-resolution Tm data processing step (3000) and the well count calculation step (3200) can be linked together to process and visualize data in real time.

[0345] Preferably, the Tm data extracted in the high-resolution Tm data processing step (3000) can be directly transferred to the well count calculation step (3200) and converted into a histogram without delay. Preferably, the ultra-precision temperature resolution-based digital melting curve analysis and visualization method can support a flexible data input format to be applicable to data generated from various PCR platforms.

[0346] Referring to FIG. 28, FIG. 28 is a conceptual flowchart of a high-resolution Tm data processing step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include a high-resolution Tm data processing step (3000) and a PCR control peak alignment step (3300).

[0347] Preferably, the high-resolution Tm data processing step (3000) can precisely extract interpolated Tm data. Preferably, the high-resolution Tm data processing step (3000) can extract Tm data by analyzing the change in fluorescence signal according to temperature change from raw data acquired from a real-time PCR instrument.

[0348] Preferably, the high-resolution Tm data processing step (3000) can accurately identify the peaks of the melting curve by calculating the first derivative of the fluorescence signal according to the temperature change. Preferably, the high-resolution Tm data processing step (3000) can detect even minute Tm differences by processing data with a temperature resolution of 0.1°C or less. Preferably, the high-resolution Tm data processing step (3000) may include a signal processing algorithm to remove background noise and improve the signal-to-noise ratio. Preferably, according to one embodiment of the present invention, the high-resolution Tm data processing step (3000) may include a PCR control peak reference alignment step (3300). Preferably, the PCR control peak reference alignment step (3300) sets the PCR control peak used in the melting analysis as a reference value and reference calibrates the Tm value of the entire sample based on this, thereby reducing errors between experiments or equipment and ensuring quantitative accuracy for comparison between data. Preferably, the PCR control peak reference alignment step (3300) can set the melting peak temperature of the standard PCR control sample included in each experiment set as the reference point.

[0349] Preferably, the PCR control peak reference alignment step (3300) can normalize the Tm data of all samples by calculating the relative temperature difference from the reference point. Preferably, the PCR control peak reference alignment step (3300) can provide a basis for integrating and analyzing data obtained from different experimental conditions or different PCR equipment.

[0350] Preferably, the PCR control peak reference alignment step (3300) can detect systemic errors that may occur during the experiment by monitoring positional changes of the PCR control peak.

[0351] Preferably, the PCR control peak reference alignment step (3300) can provide a function to accurately compare minute Tm differences between samples based on aligned data. Preferably, the PCR control peak reference alignment step (3300) may include a statistical analysis tool to verify the accuracy and reliability of the algorithm used in the alignment process.

[0352] Preferably, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution can provide a graphic interface that visualizes aligned data, allowing the user to intuitively interpret the results.

[0353] Preferably, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution can establish an automated analysis system by database-ing characteristic melting patterns for various gene variant types.

[0354] Referring to FIG. 29, FIG. 29 is a conceptual flowchart of a digital melting curve analysis and visualization method based on ultra-precision temperature resolution according to an example of an embodiment of the present invention. According to an embodiment of the present invention, the digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include an analysis software execution step (4000).

[0355] Preferably, the analysis software execution step (4000) may be a step of executing a high-resolution correction algorithm and a visualization UI in the analysis software.

[0356] Preferably, the analysis software execution step (4000) can perform the function of improving temperature resolution by processing melting curve data collected in real time.

[0357] Preferably, the high-resolution correction algorithm used in the analysis software execution step (4000) can provide a temperature resolution that is at least twice as high as the existing temperature measurement data.

[0358] Preferably, the analysis software execution step (4000) may include noise removal and signal amplification processes for the collected raw data. Preferably, the analysis software execution step (4000) may detect minute changes in the melting curve to precisely analyze changes in the nucleic acid structure.

[0359] Preferably, the visualization UI provided in the analysis software execution step (4000) may include a function that allows the user to zoom in on a specific section of the melting curve to observe detailed temperature changes. Preferably, the analysis software execution step (4000) may provide a function that allows simultaneous comparative analysis of the melting curves of various samples.

[0360] Preferably, the analysis software execution step (4000) can automatically detect inflection points of the melting curve to identify important transition temperatures. Preferably, the analysis software execution step (4000) can analyze the characteristics of the melting curve within a user-defined temperature range and provide them as numerical values.

[0361] Preferably, the analysis software execution step (4000) may include a function to export the analysis results in various formats (e.g., CSV, PDF, image files, etc.). Preferably, the analysis software execution step (4000) may provide a function to compare the previously stored analysis results with the current analysis results.

[0362] Preferably, the analysis software execution step (4000) may include a function to store and share analysis results by linking with a cloud-based database.

[0363] Preferably, the analysis software execution step (4000) can provide analysis functions accessible differentially according to the user's permission level. Preferably, the analysis software execution step (4000) can maintain stable performance even when processing large amounts of data by optimizing system resource usage.

[0364] Referring to FIG. 30, FIG. 30 is a conceptual flowchart of an analysis software execution step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include an analysis software execution step (4000) and a data smoothing step (4100).

[0365] Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may be a step of executing a high-resolution correction algorithm and a visualization UI in the analysis software. Preferably, the analysis software execution step (4000) may provide an interface that allows a user to load and analyze digital melting curve data.

[0366] Preferably, the analysis software execution step (4000) may include a high-resolution correction algorithm to improve the precision of temperature data. Preferably, the analysis software execution step (4000) may provide a function to automatically identify and analyze characteristic points of a melting curve. Preferably, the analysis software execution step (4000) may include UI elements that allow a user to visualize the analysis results in various forms. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may include a data smoothing step (4100).

[0367] Preferably, the data smoothing step (4100) may be a step that processes the data smoothly to improve the accuracy of the analysis. Preferably, the data smoothing step (4100) may apply various smoothing algorithms such as linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression-based interpolation. Preferably, the data smoothing step (4100) may generate noise-removed melting curve data to improve the accuracy of subsequent analysis.

[0368] Preferably, the data smoothing step (4100) can simultaneously visualize the original data and the smoothed data to allow the user to compare the processing results. Preferably, the analysis software execution step (4000) and the data smoothing step (4100) can be executed sequentially or in parallel.

[0369] Preferably, the analysis software execution step (4000) can provide advanced analysis functions such as differential analysis of the melting curve, peak detection, and area calculation.

[0370] Preferably, the analysis software execution step (4000) may include a function to save and export the analysis results in various formats.

[0371] Referring to FIG. 31, FIG. 31 is a conceptual flowchart of an analysis software execution step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include an analysis software execution step (4000) and a noise removal step (4200).

[0372] Preferably, the analysis software execution step (4000) may be a step of executing a high-resolution correction algorithm and a visualization UI in the analysis software. Preferably, the analysis software execution step (4000) may provide an interface for a user to load and analyze melting curve data. Preferably, in the analysis software execution step (4000), a high-resolution correction algorithm may be implemented to improve the precision of temperature data. Preferably, this algorithm provides a more precise temperature resolution than existing temperature measurement data, enabling the detection of even minute changes in the melting curve. Preferably, the analysis software execution step (4000) may include UI elements that allow a user to visualize data in various forms. Preferably, such visualization UI may represent melting curve data in various forms, such as graphs, charts, and heatmaps. Preferably, the analysis software execution step (4000) may include a function capable of processing data and displaying results in real time. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may include a noise removal step (4200). Preferably, the noise removal step (4200) may be a step that improves data quality by removing noise occurring in the data. Preferably, in the noise removal step (4200), various filtering algorithms may be applied to remove unnecessary signal fluctuations that occurred during the measurement process. Preferably, the noise removal step (4200) may utilize advanced signal processing techniques such as a moving average filter, a Gaussian filter, or a wavelet transform. Preferably, the noise removal step (4200) may be designed to selectively remove only noise while preserving the original characteristics of the data as much as possible. Preferably, the data processed through the noise removal step (4200) may contribute to more accurate melting point identification and improved reliability of analysis results.Preferably, the noise removal step (4200) may provide parameter setting options that allow the user to adjust the noise removal intensity. Preferably, this enables noise removal optimized for various experimental environments and data characteristics. Preferably, the noise removal step (4200) may also provide a visualization tool that allows comparison of data before and after processing, so that the user can intuitively verify the noise removal effect.

[0373] Referring to FIG. 32, FIG. 32 is a conceptual flowchart of an analysis software execution step according to an example of an embodiment of the present invention. According to an embodiment of the present invention, a digital melting curve analysis and visualization method based on ultra-precision temperature resolution may include an analysis software execution step (4000) and a threshold filtering step (4300).

[0374] Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may execute a high-resolution correction algorithm and a visualization UI in the analysis software. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may provide an interface for a user to load and analyze melting curve data. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may include a high-resolution correction algorithm to improve the precision of temperature data. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may provide various graph and chart options for visual representation of melting curve data.

[0375] Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may include a threshold filtering step (4300). Preferably, according to one embodiment of the present invention, the threshold filtering step (4300) may filter data based on a set threshold.

[0376] Preferably, according to one embodiment of the present invention, the threshold filtering step (4300) can identify significant change points in the melting curve data based on a user-defined temperature threshold. Preferably, according to one embodiment of the present invention, the threshold filtering step (4300) can perform the function of removing noise and improving the quality of the data. Preferably, according to one embodiment of the present invention, the threshold filtering step (4300) can increase the accuracy of the data by applying various filtering algorithms. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) and the threshold filtering step (4300) can be interconnected to derive accurate analysis results of the melting curve data. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) can provide the function of saving and exporting the data processing results in various formats.

[0377] Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may include a function for processing and visualizing data in real time. Preferably, according to one embodiment of the present invention, the threshold filtering step (4300) may provide an interface that allows a user to dynamically adjust the threshold. Preferably, according to one embodiment of the present invention, the analysis software execution step (4000) may include a statistical analysis function of melting curve data.

[0378] The ultra-precision temperature resolution-based digital melting curve analysis according to the present invention is applicable to early diagnosis or determination of progression information for diseases such as cancer and dementia; and has biological and clinical applicability such as tracking time-series changes in cell damage and abnormal methylation patterns through repeated analysis of the same specimen or patient.

Claims

Claim 1 A digital melting curve analysis and visualization system based on ultra-precision temperature resolution, comprising: a data input module that uses raw melting data collected from a real-time digital PCR platform as input values; a temperature resolution correction module that generates additional temperature points using an interpolation method based on measured temperature intervals and converts them into a high-resolution curve; and a high-resolution Tm data processing module that precisely extracts interpolated Tm data from the high-resolution curve, wherein the temperature resolution correction module includes an interpolation processing module that generates additional temperature points using an interpolation method based on data measured at temperature intervals of 0.2°C or more and converts them into a high-resolution curve having temperature intervals of 0.1°C or less, and the high-resolution Tm data processing module includes a Tm data binning processing module that generates a histogram by binning the x-axis into temperature units of 0.1°C or less. A digital melting curve analysis and visualization system based on ultra-precision temperature resolution, comprising a well count calculation module that expresses the number of wells having a corresponding Tm on the y-axis, wherein the well count calculation module performs quantitative analysis of the concentration of a target by calculating the frequency of wells belonging to a specific Tm range, and the high-resolution Tm data processing module further comprises a PCR control peak reference alignment module that sets a PCR control peak used in the melting analysis as a reference value and reference-calibrates the Tm value of the entire sample based thereon, wherein the PCR control peak reference alignment module is configured to correct errors between experiments or between equipment by adjusting the relative position relative to the reference point when comparing data before and after alignment. Claim 2 In claim 1, the data input module comprises a real-time digital PCR platform interface module that collects data in conjunction with a real-time digital PCR platform, in a digital melting curve analysis and visualization system based on ultra-precision temperature resolution. Claim 3 delete Claim 4 In claim 1, the temperature resolution correction module comprises a derivative curve generation module that generates a first derivative (-dF / dT) curve of each well to improve the precision of Tm extraction, in a digital melting curve analysis and visualization system based on ultra-precision temperature resolution. Claim 5 delete Claim 6 delete Claim 7 A digital melting curve analysis and visualization system based on ultra-precision temperature resolution, wherein the interpolation method comprises a processing module for an interpolation method based on linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression. Claim 8 delete Claim 9 A digital melting curve analysis and visualization system based on ultra-precision temperature resolution according to claim 1, comprising an analysis software module which is a software module that executes a high-resolution correction algorithm and a visualization UI. Claim 10 delete Claim 11 In claim 9, the analysis software module comprises a noise removal module that removes noise from data to improve data quality, in a digital melting curve analysis and visualization system based on ultra-precision temperature resolution. Claim 12 In claim 9, the analysis software module comprises a threshold filter module that filters data based on a set threshold, in a digital melting curve analysis and visualization system based on ultra-precision temperature resolution. Claim 13 A method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution, comprising: a data input step for receiving raw melting data collected from a real-time digital PCR platform; a temperature resolution correction step for converting measured temperature data into high-resolution temperature data; and a high-resolution Tm data processing step for precisely extracting Tm data interpolated from the high-resolution temperature data, wherein the temperature resolution correction step includes an interpolation application step for converting data measured at temperature intervals of 0.2°C or greater into a high-resolution curve having temperature intervals of 0.1°C or less by generating additional temperature points using an interpolation method based on linear interpolation, spline interpolation, polynomial interpolation, or Gaussian process regression; and the high-resolution Tm data processing step includes a Tm data binning step for generating a Tm data histogram by binning the x-axis into temperature units smaller than the temperature units of the raw input data. A method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution, comprising a well count calculation step that expresses the number of wells having a corresponding Tm on the y-axis of a histogram, wherein the high-resolution Tm data processing step further comprises a PCR control peak reference alignment step that sets a PCR control peak used in the melting analysis as a reference value and reference calibrates the Tm value of the entire sample based on this to reduce errors between experiments or equipment and ensure quantitative accuracy for comparison between data. Claim 14 In claim 13, the above data input step includes a real-time digital PCR platform data interface step that collects data in conjunction with a real-time digital PCR platform, a method for digital melting curve analysis and visualization based on ultra-precision temperature resolution. Claim 15 delete Claim 16 In claim 13, the temperature resolution correction step comprises a derivative curve generation step that generates a first derivative (-dF / dT) curve of each well to improve the precision of Tm extraction, a method for ultra-precision temperature resolution-based digital melting curve analysis and visualization. Claim 17 delete Claim 18 delete Claim 19 delete Claim 20 A method for analyzing and visualizing a digital melting curve based on ultra-precision temperature resolution according to claim 13, comprising an analysis software execution step of executing a high-resolution correction algorithm and a visualization UI in the analysis software. Claim 21 delete Claim 22 In claim 20, the analysis software execution step includes a noise removal step that improves data quality by removing noise generated in the data, a method for digital melting curve analysis and visualization based on ultra-precision temperature resolution. Claim 23 In claim 20, the analysis software execution step comprises a threshold filtering step for filtering data based on a set threshold, a method for ultra-precision temperature resolution-based digital melting curve analysis and visualization. Claim 24 In claim 1, the system is a digital melting curve analysis and visualization system based on ultra-precision temperature resolution, intended for early diagnosis or determination of progression information regarding diseases such as cancer and dementia. Claim 25 In claim 1, the system is an ultra-precision temperature resolution-based digital melting curve analysis and visualization system for tracking time-series changes in cell damage and abnormal methylation patterns through repeated analysis of the same sample or patient.

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