Method and apparatus for generating assay component

Abstract

A method for generating an assay component, comprising the following steps of: providing a plurality of processing / analysis modules based on the type of assay; selecting any one processing / analysis module from among the plurality of processing / analysis modules; setting a parameter value to be applied to each sub-module of the selected processing / analysis module; storing, in analysis information, first analysis conditions including the processing / analysis module, each sub-module, and the parameter value; additionally selecting any one processing / analysis module from among the plurality of processing / analysis modules; setting a parameter value to be applied to each sub-module of the additionally selected processing / analysis module; and additionally storing, in the analysis information, second analysis conditions including the processing / analysis module, each sub-module, and the parameter value.

Description

Method and device for creating ASSAY components

[0001] The present disclosure relates to a method and device for generating an assay component for a molecular diagnostic assay for a plurality of target nucleic acid molecules.

[0002]

[0003] Molecular diagnostics is a rapidly growing field in the in vitro diagnostics market for early disease detection. Among these, methods utilizing nucleic acids, based on their high specificity and sensitivity, are being used to diagnose genetic factors responsible for viral and bacterial infections.

[0004] Most diagnostic methods using nucleic acids utilize nucleic acid amplification reactions to amplify target nucleic acids (e.g., viral or bacterial nucleic acids). A representative example of a nucleic acid amplification reaction is the polymerase chain reaction (PCR), which involves repeated cycles of denaturation of double-stranded DNA, annealing of oligonucleotide primers to a DNA template, and extension of the primers by DNA polymerase (Mullis et al., U.S. Patent Nos. 4,683,195, 4,683,202, and 4,800,159; Saiki et al., Science 230:1350-1354, 1985). Other methods for amplifying nucleic acids include ligase chain reaction (LCR) (U.S. Pat. Nos. 4,683,195 and 4,683,202; PCR Protocols: A Guide to Methods and Applications (Innis et al., eds, 1990)), strand displacement amplification (SDA) (Walker, et al. Nucleic Acids Res. 20(7):1691-6 (1992); Walker PCR Methods Appl 3(1):1-6 (1993)), and transcription-mediated amplification (Phyffer, et al., J. Clin. Microbiol. 34:834-841 (1996); Vuorinen, et al., J. Clin. Microbiol. 33:1856-1859 (1995)). Nucleic acid sequence-based amplification (NASBA) (Compton, Nature 350(6313):91-2 (1991)), rolling circle amplification (RCA) (Lisby, Mol.These may include Q-beta replicase (Lizardi et al., BiolTechnology 6:1197(1988)), loop-mediated isothermal amplication (LAMP, Y. Mori, H. Kanda and T. Notomi, J. Infect. Chemother., 2013, 19, 404-411), recombinase polymerase amplication (RPA, J. Li, J. Macdonald and F. von Stetten, Analyst, 2018, 144, 31-67), etc.

[0005] Recently, multiplex diagnostic technologies based on these nucleic acid amplification reactions have been used to detect multiple target nucleic acids within a single tube. For example, various multiplex technologies exist for detecting multiple types of viruses simultaneously, using methods such as PCR and LAMP, as examples of nucleic acid amplification reactions.

[0006] These nucleic acid amplification reaction technologies require the use of target nucleic acid detection reagents, including oligonucleotides (e.g., primers and / or probes) that specifically hybridize to the target nucleic acid of interest, labels, DNA polymerase, dNTPs, Mg ions, and buffers, to amplify and detect the target nucleic acid of interest.

[0007] Reagents for target nucleic acid detection are developed according to molecular diagnostic development protocols. During the development process, selection of sophisticated oligonucleotides that can be developed and commercialized and various performance tests are essential.

[0008] In these performance experiments, amplification reaction result data is obtained, and as an analysis method to determine whether amplification of a meaningful target nucleic acid molecule exists in the obtained amplification reaction result data, a module is required to determine, process, and analyze the Ct (Cycle threshold) value for each sample and the signal value (e.g., RFU value) at a specific cycle in analyzing the data obtained from the reaction well of the plate as a criterion for determining presence or absence.

[0009]

[0010] The problem to be solved according to one embodiment is to provide a technology for more easily and quickly determining the values ​​of processing / analysis modules and parameters that have a major influence on the presence / rejection result in the development of a reagent (assay) for detecting a target nucleic acid molecule.

[0011] The problem to be solved according to one embodiment is to provide an interface technology that makes it easy to check and compare analysis results (presence / failure reading results) or changes (amplification curves) by changing the values ​​of processing / analysis modules and parameters in various ways.

[0012] Specifically, the Assay component generation device (100) according to the present disclosure can generate analysis conditions in various ways to simultaneously confirm or compare analysis results (presence / absence reading results) or changes (amplification curves) by changing the values ​​of processing / analysis modules and parameters that have a major influence on the presence / absence reading results, and can compare and confirm the values ​​of processing / analysis modules and / or parameters for each analysis condition.

[0013] However, the problems to be solved by the present invention are not limited to those mentioned above, and other problems to be solved that are not mentioned can be clearly understood by a person having ordinary skill in the art to which the present disclosure pertains from the description below.

[0014]

[0015] In one embodiment, a computer-implemented method for generating an Assay component used in an analysis of a performance experiment required for developing an Assay for detecting a plurality of target nucleic acid molecules is provided through an Assay component generating device including a processor and a memory operably coupled thereto, wherein the memory stores instructions executable by the processor, and the Assay component generating method comprises the following steps: providing a plurality of processing / analysis modules based on the type of the Assay; the processing / analysis modules include sub-modules for processing and analyzing amplification reaction result data of the performance experiment, and wherein any one of the plurality of processing / analysis modules is selected; setting a value of a parameter to be applied to each sub-module of the selected processing / analysis module; storing a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter in analysis information; additionally selecting any one of the plurality of processing / analysis modules; setting a value of a parameter to be applied to each sub-module of the additionally selected processing / analysis module; A step of additionally storing a second analysis condition including the processing / analysis module, each of the sub-modules, and the values ​​of the parameters in the analysis information; a step of selecting a final analysis condition from among the first and second analysis conditions stored in the analysis information; and a step of generating an Assay component using the final analysis condition.

[0016] In addition, the method is characterized in that the value of the set parameter is a value of a preset parameter for each sub-module parameter or a value of a change in the value of the preset parameter is input and set as a parameter value.

[0017] In addition, the method further includes a step of displaying the value of the preset parameter for each parameter of the sub-module when the value of the preset parameter is used as the value of the parameter of the analysis condition stored in the analysis information; or a step of receiving the change value when the change value must be input.

[0018] In addition, the method further includes a process of processing and analyzing the amplification reaction result data using the first analysis condition and displaying the analysis result; and a process of processing and analyzing the amplification reaction result data using the second analysis condition and displaying the analysis result.

[0019] In addition, the method further includes a process in which the analysis result for the first analysis condition is displayed as the correct data result of the amplification reaction result data; and a process in which the analysis result for the second analysis condition is displayed as the correct data result of the amplification reaction result data.

[0020] In addition, the method includes the amplification reaction result data for the reaction plate used in the performance experiment, the amplification reaction result data analyzed under the analysis conditions, the raw data for the amplification reaction result data, the corrected data from which the baseline is subtracted to remove the background signal value from the raw data, and the label data result indicating the presence or absence of the target for each of the plurality of target nucleic acid molecules for the performance experiment.

[0021] In addition, the method further includes a step of selecting a reaction vessel type included in the reaction plate used in the performance experiment; wherein each of the first and second analysis conditions includes values ​​of different parameters for different reaction vessel types.

[0022] In addition, the method is characterized in that each analysis condition of the stored analysis information has a different value of the parameter when the selected processing / analysis module and the additionally selected processing / analysis module are the same.

[0023] In addition, the method is characterized in that the selected analysis condition is one of the analysis conditions applied to the amplification reaction result data.

[0024] In addition, the method comprises a step of providing a section for displaying the analysis information; the section for displaying the analysis information displays an analysis condition selected from among the analysis conditions, an experimental equipment used in the performance experiment, an amplification reaction result data obtained by performing an amplification reaction on a reaction plate by the experimental equipment, information on target nucleic acid molecules for each detection channel for the reaction plate, and correct answer data results of the amplification reaction result data, and the analysis condition and the analysis result for the analysis condition are displayed together in the section for displaying the analysis information.

[0025] In addition, the method is characterized in that, in the section for displaying the analysis information, a list of at least one analysis condition is displayed, and analysis conditions and analysis results for at least one analysis condition selected from the list are displayed.

[0026] In addition, the method is characterized in that each analysis condition in the list is copyable, and by calling any one of the copied analysis conditions, all or part of the analysis conditions are modified to create another analysis condition.

[0027] In addition, the method, the performance experiment includes a plurality of performance experiments,

[0028] The method further includes a step of analyzing target amplification reaction result data by calling the selected analysis conditions for each performance experiment; wherein the selected analysis conditions are applied to each of the plurality of performance experiments.

[0029] In addition, the method is characterized by further including a step of storing the amplification reaction result data of the performance experiment as metadata for the target Assay; and a step of matching the analysis information and the metadata.

[0030] In addition, the method is characterized in that the step of providing the processing / analysis module is received as a processing / analysis component by combining a library file of one processing / analysis module selected from a plurality of processing / analysis modules provided through a processing / analysis component providing device that generates the processing / analysis module and a custom code for each assay type.

[0031] In addition, the method is characterized in that the step of displaying the analysis results is performed by providing the first and / or second analysis conditions to a calculation server that calculates the analysis conditions, performing calculations for the processing and analysis in the calculation server, and returning the first analysis result and the second analysis result for the first and / or second analysis conditions to the device.

[0032] In addition, the method is characterized in that the step of generating the Assay component includes: a step of providing a screen for inputting data for the Assay component generated in Assay information describing the plurality of target nucleic acid molecules for the Assay component; a step of displaying at least one analysis condition stored and / or selected in the Assay; and a step of generating, using the analysis conditions for each reaction vessel type included in the reaction plate used in the performance experiment based on the Assay among the displayed analysis conditions.

[0033] In addition, the method is characterized in that the Assay component includes at least one Assay, and each Assay includes a processing / analysis component including the processing / analysis module and the values ​​of the parameters and an analysis condition including an amplification reaction protocol of the performance experiment, wherein the analysis condition exists for each type of reaction vessel used in the performance experiment.

[0034] In addition, the method is characterized in that the processing / analysis module is a common module for at least two target nucleic acid molecules among the plurality of target nucleic acid molecules.

[0035] In addition, in the above method, when the processing / analysis module includes two or more sub-modules, the Assay component further includes Assay information in which the execution order of the sub-modules for processing and analyzing is predefined.

[0036] In addition, the method is characterized in that the execution order for the processing and analysis is predefined for each of the plurality of target nucleic acid molecules.

[0037] In addition, the method is characterized in that the processing / analysis module and the values ​​of the parameters are designed through a performance optimization process of the assay.

[0038] In addition, the method is characterized in that the process of optimizing the performance of the Assay includes a step of repeating the determination of values ​​of parameters referenced by pre-designated sub-modules until the performance of the Assay satisfies a predetermined standard; and if the performance of the Assay does not satisfy the predetermined standard after the repetition is performed, redesign of the pre-designated sub-modules is further performed.

[0039] In addition, the method includes, in the process of redesigning the above-described sub-modules,

[0040] It is characterized in that at least one of the following is performed: a process of modifying one or more sub-modules included in the above-mentioned pre-designated sub-modules, a process of adding sub-modules not included in the above-mentioned pre-designated sub-modules to the Assay component, and a process of deleting at least one of the above-mentioned pre-designated sub-modules if they include two or more sub-modules or changing the execution order of the above-mentioned sub-modules.

[0041] In addition, the method is characterized in that it further includes a step in which an nth analysis condition in which a value of a parameter driven by the sub-module is set is further stored in the analysis information.

[0042] According to one embodiment, a method of using a memory, a processor, and one or more programs stored in the memory and configured to be executed by the processor, wherein the at least one instruction is executed by the processor, the method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development of a plurality of target nucleic acid molecule Assays, the method comprising the steps of: providing a plurality of processing / analysis modules based on the type of the Assay, the method being performed by a computer device; The processing / analysis module includes sub-modules for processing and analyzing the amplification reaction result data of the performance experiment, and one of the plurality of processing / analysis modules is selected, a value of a parameter to be applied to each sub-module of the selected processing / analysis module is set, a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in analysis information, and one of the plurality of processing / analysis modules is additionally selected, a value of a parameter to be applied to each sub-module of the additionally selected processing / analysis module is set, a second analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in the analysis information, and a computer device that generates the selected analysis condition among the analysis conditions stored in the analysis information as an Assay component.

[0043] A computer-readable recording medium storing a computer program according to one embodiment, wherein when the computer program is executed by one or more processors included in a computer device, a method of using a memory, a processor, and one or more programs stored in the memory and configured to be executed by the processor, the method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development of a plurality of target nucleic acid molecule Assays, the method comprising: providing a plurality of processing / analysis modules based on the type of the Assay; wherein the processing / analysis modules include sub-modules for processing and analyzing amplification reaction result data of the performance experiment; wherein a processing / analysis module among the plurality of processing / analysis modules is selected; wherein a value of a parameter to be applied to each sub-module of the selected processing / analysis module is set, and a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in analysis information; wherein a processing / analysis module among the plurality of processing / analysis modules is additionally selected; A computer program that is performed including a step of setting values ​​of parameters to be applied to each sub-module of the additionally selected processing / analysis module, and storing a second analysis condition including the processing / analysis module, each of the sub-modules, and the values ​​of the parameters in the analysis information; a step of generating an analysis condition selected from among the analysis conditions stored in the analysis information as an Assay component;

[0044] A computer-readable recording medium storing a computer program according to one embodiment, wherein the computer program is executed by one or more processors included in a computer device, and a method using a memory, a processor, and one or more programs stored in the memory and configured to be executed by the processor, the method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development of a plurality of target nucleic acid molecule Assays, the method comprising: providing a plurality of processing / analysis modules based on the type of the Assay; wherein the processing / analysis modules include sub-modules for processing and analyzing amplification reaction result data of the performance experiment; wherein any one of the plurality of processing / analysis modules is selected; wherein the value of a parameter to be applied to each sub-module of the selected processing / analysis module is set, and a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in analysis information; wherein any one of the plurality of processing / analysis modules is additionally selected; A computer-readable recording medium that is performed including a step of setting values ​​of parameters to be applied to each sub-module of the additionally selected processing / analysis module, storing a second analysis condition including the processing / analysis module, each of the sub-modules, and the values ​​of the parameters in the analysis information; and a step of generating an analysis condition selected from among the analysis conditions stored in the analysis information as an Assay component.

[0045]

[0046] According to one embodiment, a researcher and developer of a reagent for determining the presence or absence of a target nucleic acid molecule in a sample can be provided with an optimal processing / analysis module applicable to the process of determining the presence or absence (positive or negative).

[0047] In addition, you can receive default values ​​(values ​​of preset parameters) for parameters based on the processing / analysis module as a reference, or input new values ​​by changing the default values ​​when necessary, and perform analysis on the amplification reaction result data based on these, and immediately check the analysis results.

[0048] In addition, by performing analysis by creating various analysis conditions, it is possible to compare / contrast the presence / failure correct answers for various analysis conditions.

[0049] In addition, the developer can quickly and accurately set the values ​​of the processing / analysis module and parameters so that the analysis results are as identical as possible to the correct answer of the presence or absence of the amplification reaction result data by processing / analyzing the amplification reaction result data while changing the analysis conditions in various ways.

[0050] Additionally, by selecting the most appropriate analysis conditions according to the Assay type and packaging them into the Assay component, the processing / analysis modules and parameter values ​​can be efficiently distributed and managed.

[0051] Through this, in molecular diagnosis using assays, it is possible to easily manage which processing / analysis module should be applied to process or analyze amplification data, and which parameter should be referenced as which value at this time.

[0052]

[0053] FIG. 1 illustrates an assay component generation device and a device connected thereto according to one embodiment.

[0054] FIG. 2 is a block diagram of an assay component generation device according to one embodiment.

[0055] Figures 3 to 5 and Figure 7 illustrate implementation examples of interface provision screens in an Assay component generation device according to one embodiment.

[0056] Figure 6 is a flowchart illustrating a method for creating an Assay component according to one embodiment.

[0057]

[0058] The advantages and features of the present invention, and the methods for achieving them, will become clearer with reference to the embodiments described in detail below together with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below and may be implemented in various different forms. These embodiments are provided solely to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention, and the present invention is defined solely by the scope of the claims.

[0059] When describing embodiments of the present invention, detailed descriptions of known functions or configurations will be omitted if they are deemed to unnecessarily obscure the gist of the invention. Furthermore, the terms described below are defined in light of their functions in the embodiments of the present invention and may vary depending on the intent or custom of the user or operator. Therefore, their definitions should be based on the overall content of this specification.

[0060] Before explaining Figure 1, let us look at the terms used herein.

[0061] The term "target analyte" encompasses a variety of substances (e.g., biological and non-biological substances), which may refer to the same entity as the term "target analyte."

[0062] Such target analytes may specifically include biological materials, more specifically at least one of nucleic acid molecules (e.g., DNA and RNA), proteins, peptides, carbohydrates, lipids, amino acids, biological compounds, hormones, antibodies, antigens, metabolites, and cells.

[0063] The term "sample" refers to biological samples (e.g., cells, tissues, and body fluids) and non-biological samples (e.g., food, water, and soil). Among these, the biological samples may include at least one of, for example, viruses, bacteria, tissues, cells, blood (including whole blood, plasma, and serum), lymph, bone marrow fluid, saliva, sputum, swabs, aspirations, milk, urine, stool, eye fluid, semen, brain extracts, spinal fluid, joint fluid, thymus fluid, bronchial lavage fluid, ascites, and amniotic fluid. These samples may or may not contain the aforementioned target analytes.

[0064] Meanwhile, if the target analyte described above is a nucleic acid molecule or contains a nucleic acid molecule, a nucleic acid extraction process known in the art may be performed on the sample presumed to contain the target analyte (see: Sambrook, J. et al., Molecular Cloning. A Laboratory Manual, 3 rd ed. Cold Spring Harbor Press (2001)). The nucleic acid extraction process may vary depending on the type of sample. In addition, if the extracted nucleic acid is RNA, a reverse transcription process may be additionally performed to synthesize cDNA (Reference: Sambrook, J. et al., Molecular Cloning. A Laboratory Manual, 3rd ed. Cold Spring Harbor Press (2001)).

[0065] The term "data set" refers to data obtained from a signal generation response for the target analyte using a signal generation means (the signal generation means will be described later).

[0066] In this case, the term "signal generating reaction" refers to a reaction that generates a signal dependent on the properties of a target analyte in a sample, such as activity, amount, or presence (or absence), specifically the presence (or absence). Such signal generating reactions include biological reactions and chemical reactions. Among these, biological reactions include genetic analysis processes such as PCR, real-time PCR, isothermal amplification, and microarray analysis, immunological analysis processes, and bacterial growth analysis. In addition, chemical reactions include processes that analyze the production, change, or destruction of a chemical substance. According to one embodiment, the signal generating reaction may be a genetic analysis process, or may be a nucleic acid amplification reaction, an enzymatic reaction, or microbial growth.

[0067] Meanwhile, the aforementioned signal generation response is accompanied by a signal change. Therefore, the progress of this signal generation response can be assessed by measuring the signal change.

[0068] Here, the term "signal" refers to a measurable output. Furthermore, the measured magnitude or change in this signal serves as an indicator, either qualitatively or quantitatively, of the characteristics of the target analyte, specifically the presence or absence of the target analyte in the sample.

[0069] Here, examples of indicators include, but are not limited to, fluorescence intensity, luminescence intensity, chemiluminescence intensity, bioluminescence intensity, phosphorescence intensity, charge transfer, voltage, current, power, energy, temperature, viscosity, light scatter, radiation intensity, reflectance, transmittance, and absorbance.

[0070] The term "signal generating means" as mentioned above means a means for providing a signal indicating the characteristics, specifically the presence or absence, of the target analyte to be analyzed.

[0071] Such signal generating means include the label itself or an oligonucleotide to which the label is linked.

[0072] Among these, the labels include fluorescent labels, luminescent labels, chemiluminescent labels, electrochemical labels, and metal labels. The labels may be used as labels themselves, such as intercalating dyes. Alternatively, the labels may be used in the form of a single label or an interactive dual label comprising a donor molecule and an acceptor molecule, bound to one or more oligonucleotides.

[0073] When using a fluorescent label, the signal value can be expressed as a RFU (Relative Fluorescence Unit) value.

[0074] The signal generating means may additionally include an enzyme having a nucleic acid cleavage activity to generate a signal (e.g., an enzyme having a 5' nucleic acid cleavage activity or an enzyme having a 3' nucleic acid cleavage activity).

[0075] Meanwhile, various methods for generating a signal indicating the presence of a target analyte, particularly a target nucleic acid molecule, using the above signal generating means are known. Representative examples include: TaqMan TMProbe method (US Patent No. 5,210,015), molecular beacon method (Tyagi, Nature Biotechnology, v.14 MARCH 1996), Scorpion method (Whitcombe et al., Nature Biotechnology 17:804-807 (1999)), Sunrise (or Amplifluor) method (Nazarenko et al., Nucleic Acids Research, 25(12):2516-2521 (1997), and US Patent No. 6,117,635), Lux method (US Patent No. 7,537,886), CPT (Duck P, et al. Biotechniques, 9:142-148 (1990)), LNA method (US Patent No. 6,977,295), Plexor method (Sherrill CB, et al., Journal of the American Chemical Society, 126:4550-4556(2004)), Hybeacons (DJ French, et al., Molecular and Cellular Probes 13:363-374(2001) and U.S. Pat. No. 7,348,141), dual-labeled, self-quenched probes (U.S. Pat. No. 5,876,930), hybridization probes (Bernard PS, et al., Clin Chem 2000, 46, 147-148), PTOCE (PTO cleavage and extension) method (WO 2012 / 096523), PCE-SH (PTO Cleavage and Extension-Dependent Signaling Oligonucleotide Hybridization) method (WO2013 / 115442), PCE-NH (PTO Cleavage and Extension-Dependent Non-Hybridization) method (PCT / KR2013 / 012312) and CER method (WO 2011 / 037306).

[0076] Meanwhile, the aforementioned term "signal generation reaction" may include a signal amplification reaction. In this case, the term "amplification reaction" refers to a reaction that increases or decreases a signal generated by the signal generation means. Specifically, the amplification reaction refers to a reaction that increases (or amplifies) a signal generated by the signal generation means depending on the presence of a target analyte.

[0077] In such amplification reactions, amplification of a target analyte (e.g., a nucleic acid molecule) may or may not be accompanied. More specifically, the amplification reaction may refer to a signal amplification reaction accompanied by amplification of a target analyte.

[0078] Meanwhile, the data set obtained through the above amplification reaction may include an amplification cycle.

[0079] Here, the term "cycle" refers to a unit of change in a condition in a plurality of measurements involving a change in said condition. The change in said constant condition may refer to an increase or decrease in, for example, temperature, reaction time, number of reactions, concentration, pH, or the number of replications of the measurement target (e.g., nucleic acid). Accordingly, a cycle may be a time or process cycle, a unit operation cycle, or a reproductive cycle.

[0080] More specifically, the term "cycle" means one unit of repetition, when a reaction is repeated in a certain process or at certain time intervals.

[0081] Alternatively, the term "cycle" may mean one unit of repetition when a certain action is repeated as the reaction progresses.

[0082] For example, when a nucleic acid amplification reaction is performed, the act of detecting a signal generated at regular time intervals may be repeated, and may represent one unit of said repetition. In this case, a cycle may have a unit of time.

[0083] For example, in the case of a nucleic acid amplification reaction, one cycle refers to a reaction that includes the steps of nucleic acid denaturation, primer annealing, and primer extension. In this case, a change in a certain condition is an increase in the number of reaction repetitions, and the unit of repetition of the reaction comprising the above series of steps is defined as one cycle. The number of cycles may include the number of reactions or the reaction time.

[0084] Meanwhile, the aforementioned target analytes or target analytes, especially target nucleic acid molecules, can be amplified by various methods: polymerase chain reaction (PCR), ligase chain reaction (LCR) (U.S. Patent Nos. 4,683,195 and 4,683,202; PCR Protocols: A Guide to Methods and Applications (Innis et al., eds, 1990)), strand displacement amplification (SDA) (Walker, et al. Nucleic Acids Res. 20(7):1691-6 (1992); Walker PCR Methods Appl 3(1):1-6 (1993)), transcription-mediated amplification (Phyffer, et al., J. Clin. Microbiol. 34:834-841 (1996); Vuorinen, et al., J. Clin. Microbiol. 33:1856-1859 (1995)), nucleic acid sequence-based amplification (NASBA) (Compton, Nature 350(6313):91-2 (1991)), rolling circle amplification (RCA) (Lisby, Mol. Biotechnol. 12(1):75-99 (1999); Hatchet et al., Genet. Anal. 15(2):35-40 (1999)) and Q-Beta Replicase (Lizardi et al., BiolTechnology 6:1197(1988)), Loop-mediated isothermal amplication (LAMP, Y. Mori, H. Kanda and T. Notomi, J. Infect. Chemother., 2013, 19, 404-411), Recombinase polymerase amplication (RPA, J. Li, J. Macdonald and F. von Stetten, Analyst, 2018, 144, 31-67).

[0085] Meanwhile, an amplification reaction amplifies the signal while amplifying the target analyte (specifically, the target nucleic acid molecule). For example, the amplification reaction is performed according to PCR, specifically real-time PCR, or an isothermal amplification reaction (e.g., LAMP or RPA).

[0086] Meanwhile, the data set obtained by the signal generation reaction includes a plurality of data points including cycles of the signal generation reaction and signal values ​​in the cycles.

[0087] Here, the term "signal value" means a value quantified according to a certain scale of the level of a signal (e.g., signal intensity) actually measured in a cycle of a signal generation reaction, particularly an amplification reaction, or a modified value thereof. The modified value may include a mathematically processed signal value of the actually measured signal value. Examples of mathematically processed signal values ​​of the actually measured signal value (i.e., the signal value of raw data) may include logarithmic values ​​or derivatives.

[0088] The term "data point" refers to a single coordinate value that includes a cycle and a signal value. Furthermore, the term "data" refers to all information that constitutes a data set. For example, each cycle and signal value of an amplification reaction can be considered data.

[0089] Data points obtained by a signal generation reaction, particularly an amplification reaction, can be expressed as coordinate values ​​that can be represented in a two-dimensional rectangular coordinate system. In the coordinate values, the X-axis represents the corresponding cycle number, and the Y-axis represents the signal value measured or processed in the corresponding cycle.

[0090] The term "data set" refers to a collection of data points. For example, the data set may be a collection of data points obtained directly through an amplification reaction performed in the presence of a signal generation means, or may be a modified data set obtained by modifying such a data set. The data set may be a portion or all of a plurality of data points obtained through the amplification reaction, or a modified data set thereof.

[0091] Meanwhile, the data set may be obtained by processing multiple data sets. When analyzing multiple target analytes in a single reaction vessel, the data set for the multiple target analytes may be obtained by processing the data sets obtained from the reactions conducted in the single reaction vessel. For example, the data set for multiple target analytes in a single reaction vessel may be obtained by processing multiple data sets obtained from signals measured at different temperatures.

[0092] The aforementioned data set can be plotted, thereby obtaining an amplification curve.

[0093] The term "reaction plate" refers to the basic unit where the amplification reaction is performed in an amplification device, and the basic unit where the data generated after the amplification reaction is stored. Different plates may be plates where the amplification reaction was performed at different times using the same amplification device, or plates where the amplification reaction was performed at the same time by different amplification devices.

[0094] A reaction plate comprises a plurality of reaction wells. The plate may comprise N x M reaction wells. Typically, the plate comprises 12 x 8 or 8 x 12 reaction wells. The reaction wells of the plate may be integral with the plate or may be in the form of detachable tubes. The plate may be rectangular in shape, but the plate may also be implemented in various shapes, such as circular, ladder-shaped, and diamond-shaped, as long as it comprises one or more reaction wells.

[0095] The wells of the plate contain the sample to be analyzed and the reagents required for the nucleic acid amplification reaction.

[0096] With reference to the drawings below, various implementation examples of the present invention will be examined.

[0097] FIG. 1 illustrates an assay component generation device (100) according to one embodiment, a nucleic acid detection device (200), a research and development data device (210), a target presence / absence reading SW device (230), and a database (220) connected thereto. These devices (100, 200, 210, 220, 230) may be connected to each other via wired or wireless communication.

[0098] However, FIG. 1 is merely exemplary, and the spirit of the present disclosure is not limited to what is illustrated in FIG. 1. For example, these (100, 200, 210, 220, 230) may further include a configuration not illustrated in FIG. 1, which may further include, but is not limited to, a server for deriving candidate nucleotides in silico, a preparation device for performing a preparation process for nucleic acid amplification, and / or a nucleic acid detection device (200), a processing / analysis component generation device for generating a processing / analysis module described below, and a calculation server device for performing calculations based on the processing / analysis module.

[0099] A result analysis module may be included here to determine whether a specific target analyte is positive or negative based on the detected result or to provide the detected result as a graph.

[0100] Meanwhile, unlike what is shown, the Assay component generation device (100) may be implemented by being included in the research and development data management device (210). However, the following description will assume that the aforementioned components (100, 200, 210, 220, 230) are implemented or connected as shown in Fig. 1. Each component will be examined in detail below.

[0101] The nucleic acid detection device (200) is implemented to perform a nucleic acid amplification reaction and a nucleic acid detection operation on a sample. Depending on the embodiment, the nucleic acid detection device (200) may be implemented to perform a nucleic acid detection operation without performing a nucleic acid amplification reaction. However, the following description will assume that the nucleic acid detection device (200) is implemented to also perform a nucleic acid amplification reaction.

[0102] Meanwhile, as the above-described nucleic acid amplification reaction is performed, if the sample in the reaction vessel contains a target nucleic acid molecule, not only will its amount be amplified, but the magnitude of the signal generated by the above-described signal generating means may also be amplified as described above. Accordingly, the nucleic acid detection device (200) is implemented to detect the magnitude of such a signal. Specifically, the nucleic acid detection device (200) can monitor in real time the magnitude of the signal that changes according to the progress of the nucleic acid amplification reaction. The monitored result can be output from the nucleic acid detection device (200) in the form of a data set as mentioned in the definition of terms, for example, the intensity of the signal per cycle. Here, the intensity of the signal per cycle is information that serves as a basis for determining the presence or absence of the target nucleic acid molecule in the corresponding sample.

[0103] Meanwhile, since the size of the signal generated by the signal generating means can also be amplified when the above nucleic acid amplification reaction is performed, the nucleic acid amplification reaction will be considered hereinafter as causing the signal generating reaction discussed above as a term.

[0104] Various types of data are stored in the database (220).

[0105] For example, data regarding the aforementioned nucleic acid amplification reaction may be stored in the database (220). Specifically, the signal intensity for each of multiple cycles for amplifying nucleic acids, the shape of the amplification curve generated using the signal intensity for each cycle, or the positive / negative reading results may be stored as research and development data. In the present specification, such "data regarding the nucleic acid amplification reaction" may include amplification reaction result data.

[0106] In addition, meta-data regarding such research and development data may be stored in the database (220). Meta-data may be described as "data about data," i.e., data that provides information about other data. For example, at least one of information regarding the oligonucleotide itself, the type of performance verification experiment to be performed on the oligonucleotide, the materials used in the experiment, the experimental procedure, and the experimental equipment for the experiment may be included in such metadata. The aforementioned items that may be included in the metadata will be discussed in more detail below.

[0107] Additionally, in the present disclosure, the oligonucleotide may include a plurality of candidate oligonucleotides, and may include an oligonucleotide selected from the plurality of candidate oligonucleotides and ultimately included in a reagent for detecting a target nucleic acid molecule.

[0108] The assay component generation device (100) can process / analyze amplification reaction result data according to the performance test results to obtain analysis results.

[0109] Performance testing is a test for the development of a reagent for detecting a specific target nucleic acid molecule, and consists of multiple tests, each of which is different from the others.

[0110] Each of these multiple performance experiments is a performance optimization experiment that selects an oligonucleotide to be included in a reagent for detecting the target nucleic acid molecule from a plurality of candidate oligonucleotides and optimizes the performance of the selected oligonucleotide.

[0111] In the present disclosure, a performance experiment means one of a plurality of performance experiments, and analysis information, analysis conditions, and analysis results generated based on the performance experiments described below can all be generated from each of the plurality of performance experiments.

[0112] The assay component generation device (100) provides a processing / analysis module used for processing / analyzing amplification reaction result data for each performance experiment among a plurality of performance experiments, sets the values ​​(set values) of parameters referenced by the processing / analysis module, analyzes the amplification reaction result data with the values ​​of the set parameters, and displays the analysis results.

[0113] Processing / analysis is performed through the processing / analysis module described below and parameter value setting. The Assay component generation device (100) according to the present disclosure may provide at least one processing / analysis module and an interface for parameter value setting for parameters based on the processing / analysis module.

[0114] In the present disclosure, the values ​​of the processing / analysis module and parameters mean analysis conditions for analyzing the amplification reaction result data of the performance experiment.

[0115] The Assay component generation device (100) according to the present disclosure can provide an interface capable of generating multiple analysis conditions so that analysis can be performed by variously changing the values ​​of processing / analysis modules and parameters in the process of analyzing amplification reaction result data for each performance experiment.

[0116] By providing this interface,

[0117] First, researchers and developers of reagents for determining the presence or absence of target nucleic acid molecules in a sample are provided with various processing / analysis modules applicable to the process of determining the presence or absence (positive or negative), thereby enabling them to select the optimal processing / analysis module.

[0118] Second, default values ​​for parameters based on the processing / analysis module can be provided as a reference, or new values ​​can be input by changing the default values ​​when necessary, and analysis of the amplification reaction result data can be performed based on these values, and the analysis results can be immediately confirmed.

[0119] Third, by creating various analysis conditions and performing analysis, it is possible to compare / contrast with the presence / failure correct answer for each analysis condition.

[0120] Fourth, the developer can quickly and accurately set the values ​​of the processing / analysis module and parameters so that the analysis results are as identical as possible to the correct answer for the presence or absence of the amplification reaction result data by processing / analyzing the amplification reaction result data while changing the analysis conditions in various ways.

[0121] Fifth, by selecting the most appropriate analysis conditions for each assay type and packaging them into the Assay component, processing / analysis modules and parameter values ​​can be efficiently distributed and managed. This facilitates management of which processing / analysis modules should be applied to process or analyze amplification reaction result data in assay-based molecular diagnostics, as well as which parameters and values ​​should be referenced.

[0122] Below, the Assay component generation device (100) according to the present disclosure will be examined in more detail.

[0123] FIG. 2 is a block diagram of an assay component generation device (100) according to one embodiment.

[0124] Referring to FIG. 2, the Assay component generation device (100) includes, but is not limited to, a communication unit (110), a memory (120), a processor (130), and a display unit (140).

[0125] First, the communication unit (110) is implemented as a wired or wireless communication module. Through this communication unit (110), the computer device (100) can communicate with the outside. For example, the assay component generation device (100) can receive amplification reaction result data from the nucleic acid detection device (200) through the communication unit (110). In addition, the assay component generation device (100) can receive information necessary for analyzing the amplification reaction result data from an external source, for example, the research and development data management device (210) and / or database (220) illustrated in FIG. 1.

[0126] The memory (120) stores various types of data or information, including at least one command. The stored data may be data received from the database (220) via the communication unit (110) or data processed by the processor (130). This data may include information on various types of analysis conditions required for generating assay components.

[0127] Meanwhile, in FIG. 2, the memory (120) is depicted as a separate configuration from the processor (130), but the memory (120) may be implemented as a single device with the processor (130). For example, the memory (120) may be a storage such as a cache included within the processor (130).

[0128] Next, the processor (130) may be implemented by a central processing unit (CPU), a graphics processing unit (GPU), a microcontroller unit (MCU), or a dedicated processor in which the methods according to one embodiment are performed. Hereinafter, such a processor (130) may collectively refer to a single processor or multiple processors, for example, a multi-core processor.

[0129] This processor (130) can write data to the memory (120). In addition, the processor (130) can read out and execute instructions stored in the memory (120). For example, the processor (130) can cause the Assay component generation device (100) to perform the functions described below by executing instructions stored in the memory (120). These functions will be described later.

[0130] The display unit (140) can be driven by the processor (130) as follows.

[0131] First, information can be displayed on the display unit (140). In addition, a user can input certain information through the display unit (140). For such display or input, the display unit (140) can be implemented as a touch screen or a touch pad, or alternatively, can be implemented in combination with an LCD monitor and a keyboard. Here, the information displayed on the display unit (140) may be transmitted from the outside to the research and development data management device (100) through the communication unit (110) or loaded from the memory (120), but is not limited thereto.

[0132] According to an embodiment, the display unit (140) displays an input screen for receiving analysis conditions. The content included in this screen may be transmitted to the Assay component creation device (100) from an external source, such as a database (220), via the communication unit (110), or loaded from the memory (120), but is not limited thereto.

[0133] Below, each functional configuration of the Assay component generation device (100) will be examined in more detail with reference to FIGS. 3 to 5.

[0134] An assay component generation device (100) according to one embodiment is used in target presence / absence reading software that displays amplification reaction data.

[0135] The device for generating a component for assay (100) is a device that obtains amplification reaction result data and generates a component for assay using the amplification reaction result data. The device receives amplification reaction result data of a performance experiment required for developing a reagent for detecting a target nucleic acid molecule from a research and development data management device (30), and processes / analyzes the received amplification reaction result data to obtain an analysis result.

[0136] To this end, the Assay component generation device (100) provides an interface used for selecting a processing / analysis module used in the process of determining the presence or absence of a target nucleic acid molecule and setting the values ​​of parameters for the selected processing / analysis module.

[0137] FIG. 3 illustrates an implementation example of an interface provision screen (300) provided by an Assay component generation device (100) according to one embodiment.

[0138] According to one embodiment, in the interface provision screen (300), a processing / analysis module for processing / analyzing the amplification reaction result data of a performance experiment is selected in section 330, and parameter value setting for the parameter based on the processing / analysis module selected in section 350 is performed.

[0139] The Assay component generation device (100) according to the present disclosure provides a plurality of processing / analysis modules based on the type of Assay through the interface provision screen (300) illustrated in FIG. 3.

[0140] Molecular diagnostic assays can be categorized based on their target nucleic acid molecules, allowing for preset processing / analysis modules. For example, processing / analysis modules can be divided into a default processing / analysis module for respiratory assays, a default processing / analysis module for intestinal bacteria assays, and a default processing / analysis module for sexually transmitted infections (STIs), allowing for specific default processing / analysis modules for each assay.

[0141] Accordingly, the interface provision screen (300) of the Assay component generation device (100) provides a preset processing / analysis module according to the corresponding Assay type without a separate input from the user for the processing / analysis module.

[0142] The default processing / analysis module for each assay type may be empirically selected based on repeated performance experiment analysis by research developers and preset in the assay component generation device (100), but is not limited thereto.

[0143] In addition, according to the present disclosure, one of the multiple processing / analysis modules provided by the Assay component generation device (100) can be selected through the interface provision screen (300) of FIG. 3. In this case, according to an embodiment, the processing / analysis module can be selected from a pre-stored processing / analysis module list (410) as illustrated in FIG. 4. At this time, the Assay component generation device (100) displays information (type, version, description, etc.) for each processing / analysis module pre-stored in the processing / analysis module selection screen (400) in a list for each processing / analysis module, thereby allowing selection of an appropriate processing / analysis module according to a performance experiment.

[0144] In addition, according to an embodiment, the Assay component creation device (100) may request the creation of a new processing / analysis module that is not provided in the interface provision screen (300) and / or the processing / analysis module list. To this end, the Assay component creation device (100) communicates with a processing / analysis component providing device (not shown) that creates a processing / analysis module, and may select a desired processing / analysis module from various processing / analysis modules provided through the processing / analysis component providing device.

[0145] The processing / analysis component providing device can generate a processing / analysis component by combining a library file of any selected processing / analysis module and a custom code for each assay type, and provide it to the assay component generating device (100).

[0146] The library file at this time refers to a file that enables the Assay component generation device (100) to be operated on the web according to an embodiment. The custom code refers to additional code for specific analysis by Assay type when executing a processing / analysis module for processing / analyzing amplification reaction result data in the Assay component generation device (100) using a commonly known javascript code.

[0147] In more detail, the processing / analysis operation server used for processing / analysis of the Assay component generation device (100) will be described later.

[0148] In one embodiment, the processing / analysis module includes a sub-module for analyzing amplification reaction result data of a performance experiment.

[0149] The processing / analysis module is a module for processing and analyzing amplification reaction result data for multiple target nucleic acid molecules in the target presence / absence reading software.

[0150] The target nucleic acid molecule presence / absence reading software is software that can confirm the presence / absence of a target nucleic acid molecule by applying the obtained amplification reaction result data to one or more reading processing / analysis modules in a predetermined order.

[0151] In one embodiment, the target presence reading software may be a program installed on a target presence reading terminal.

[0152] The target presence / absence analysis software may include a function for displaying the results of the Assay component's work. For example, the target presence / absence analysis software may be a program capable of displaying the results of processing and analysis performed by the Assay component in at least one of the following formats: numbers, charts, or graphs.

[0153] As described above, the processing / analysis module includes submodules, which in one embodiment may be algorithms or software. Among these submodules, the submodules called during the processing and analysis of amplification reaction result data may be predefined. For example, when processing and analyzing amplification reaction result data, all submodules may be called, some necessary submodules may be called, or only one submodule may be called.

[0154] The processing / analysis module may include at least one sub-module. As shown in Fig. 3, the processing / analysis module may include sub-module a (BPN), sub-module b (cut-off), and sub-module c (DSP).

[0155] The values ​​of processing / analysis modules and parameters can be designed through the Assay performance optimization process.

[0156] The assay performance optimization process can iterate through determining the values ​​of parameters referenced by predefined submodules until the assay performance meets a predetermined criterion. If the assay performance does not meet the criterion after the iteration, further redesign of the predefined submodules can be performed.

[0157] The process of redesigning the designated sub-modules may include at least one of modifying one or more sub-modules included in the designated sub-modules, adding sub-modules not included in the designated sub-modules to the Assay component, deleting at least one of the sub-modules if the designated sub-modules include two or more sub-modules, or changing the execution order of the sub-modules.

[0158] In one embodiment, it may be a common module for at least two target nucleic acid molecules among a plurality of target nucleic acid molecules.

[0159] The assay component may include common sub-modules that are commonly used for at least two of the plurality of target nucleic acid molecules. That is, the assay component may include sub-modules that are commonly used by each of the plurality of target nucleic acid molecules.

[0160] Submodules may be various modules for processing and analysis. In one embodiment, the submodules may be algorithms that analyze or mathematically process data sets obtained during the amplification process of multiple target nucleic acid molecules.

[0161] For example, a submodule may include an algorithm that outputs a cycle count when a data set obtained during the amplification process of multiple target nucleic acid molecules reaches a threshold and determines the presence or absence of the target nucleic acid molecule based at least in part on the output cycle count. Here, the cycle count may be defined as the number of reactions or time.

[0162] Additionally, a submodule may be an algorithm that performs at least one task selected from the following group.

[0163] (i) Analysis of signals detected at a relatively high detection temperature and a relatively low detection temperature during the amplification process of multiple target nucleic acid molecules;

[0164] (ii) normalizing multiple data sets obtained during the amplification process of multiple target nucleic acid molecules;

[0165] (iii) A task of determining the presence or absence of a target nucleic acid molecule using a parameter representing the maximum derivative of a sigmoid fitting curve for a data set obtained during the amplification process of multiple target nucleic acid molecules;

[0166] (iv) A task of determining the shape of a sigmoid fitting curve using a value representing the shape of a data set obtained during the amplification process of multiple target nucleic acid molecules;

[0167] (v) a task of performing regression analysis on a data set obtained during the amplification process of multiple target nucleic acid molecules; and

[0168] (vi) Performing regression model diagnosis on a portion of a data set obtained through a signal generation reaction for a target nucleic acid molecule.

[0169] The sub-module may be a sub-module for subtracting a baseline signal included in the amplification reaction result data, a sub-module for fitting the amplification reaction result data with a non-linear function to obtain a predetermined curve, and a module for deciphering the presence or absence of a target nucleic acid molecule from the obtained curve.

[0170] Specifically, the sub-module for subtracting the baseline signal included in the amplification reaction result data includes a module for obtaining corrected data by applying the normalization coefficient disclosed in WO 2017 / 086762 of the present applicant to the signal values.

[0171] The above standardization coefficient is provided using a reference value, a reference cycle, and a data set. The reference cycle is selected from the cycles of the data set, and the reference value is an arbitrarily determined value. The standardization coefficient is provided by determining the relationship between the signal value of the cycle corresponding to the reference cycle in the data set and the reference value, and the standardization coefficient is applied to the signal values ​​of the raw data to obtain corrected signal values ​​and provide them to the raw data. For example, a reference value. The reference value is a value used to provide a standardization coefficient. In the present specification, the reference value means an arbitrary value applied to the reference cycle to correct the signal values ​​of the data set. The same reference value is applied to data sets to be standardized with the same standard. If the data sets to be corrected are multiple data sets, the multiple data sets can be corrected by the same reference value. The reference value may be an arbitrarily determined value. Preferably, the reference value is an arbitrarily determined value among non-zero real numbers. The reference value may be arbitrarily selected by the experimenter as long as the presence of the target nucleic acid molecule in the sample can be determined by the correction data set using the reference value. Therefore, the reference value to be used for correction of the data set can be determined within the range of signal values ​​that can be obtained in the reference cycle by the same type of signal generation reaction as the signal generation reaction from which the data set was obtained. The reference value can be obtained separately from the data set to be corrected. Specifically, the reference value can be determined from a data set obtained from a signal generation reaction for a target nucleic acid molecule of the same type as the target nucleic acid molecule to be analyzed. Alternatively, the reference value can be obtained from a group of data sets including the data set to be corrected. Preferably, the reference value can be a value of the same type as the signal value of the data set to be corrected, and can be in the same unit or dimension as the data set to be corrected.However, even if the reference value and the signal values ​​of the data set have different units or dimensions, or the reference value has no units or dimensions, an appropriate standardization coefficient for each response can be provided from the reference value and the signal values ​​of the data set to be corrected, and the correction data set can be obtained using the standardization coefficient for each response.

[0172] Next, a sub-module that obtains a predetermined curve by fitting the amplification reaction result data with a non-linear function can perform analysis to determine the presence or absence of a target nucleic acid molecule using DSP (WO2019 / 066572) technology.

[0173] Parameters set using DSP technology include parameters used in a processed performance data set, which is obtained by mathematically processing raw data. Here, the processed performance data set encompasses not only data sets obtained by mathematically processing raw data, but also data obtained by further mathematically processing the resulting mathematically processed data. For example, data includes the first derivative of the raw data and the second, third, and other derivatives obtained from the first derivative.

[0174] The parameters of the DSP can be applied to the function for setting the value of the Ct parameter described above. For example, the DSP uses the fitting accuracy of a nonlinear function for a data set as a direct indicator for target analyte analysis, obtains a data set for a target nucleic acid molecule from a signal-generating reaction using a signal-generating means to analyze the target nucleic acid molecule without false positive and false negative results, especially false positive results, corrects the obtained data set including a plurality of data points including cycle numbers and signal values, generates a nonlinear function for the corrected data set, determines the fitting accuracy of the nonlinear function for the corrected data set, and determines the presence or absence of the target analyte in the sample using the fitting accuracy. This analysis method requires various parameters and can be optimized according to the values ​​set for each parameter. Using DSP (WO2019 / 066572) technology, the parameters used for determining the presence or absence of the target nucleic acid molecule are assigned, and the analysis is performed by setting the values ​​of the DSP parameters.

[0175] Next, the sub-module for reading the presence or absence of a target nucleic acid molecule from a curve obtained by fitting the amplification reaction result data with a non-linear function may include a module for extracting only data for each target nucleic acid from data obtained at a plurality of different temperatures for a plurality of target nucleic acids. For example, the modules disclosed in WO2015-147370, WO2015 / 147412, WO2015 / 147382, and WO2016 / 093619 of the present applicant that utilize a plurality of target detection temperatures can be used to assign parameters used in the process of reading the presence or absence of a target nucleic acid molecule, and the values ​​of the assigned parameters can be set to analyze the data of the raw data.

[0176] Even when performing analysis by setting values ​​for parameters of a technology utilizing DSP and multiple target detection temperatures, the Ct parameter can be set by adjusting the parameter values ​​through comparative analysis based on the correct answer data labeled as positive and negative, just as when performing analysis by setting the value of the Ct parameter. For example, the parameter values ​​can be adjusted so that the samples labeled as positive and negative in the correct answer data match the analysis results by setting the parameter values.

[0177] Predefined sub-modules can process and analyze amplification reaction result data for multiple target nucleic acid molecules by referencing parameter values. At this time, the pre-defined sub-modules can process and analyze the amplification reaction result data at the presence / absence detection terminal according to the request of the target presence / absence detection software. The parameter values ​​can be referenced by the sub-modules.

[0178] As described above, the Assay component generation device (100) provides a plurality of processing / analysis modules. When a selection for one of the plurality of processing / analysis modules is input through the interface provision screen (300), the values ​​of parameters to be applied to each sub-module of the selected processing / analysis module are set.

[0179] According to an embodiment, the value of the set parameter may be a value of a preset parameter for each sub-module parameter, or a value of a parameter set by inputting a change value of the value of the preset parameter.

[0180] To this end, the interface provision screen (300) can display parameters based on the selected processing / analysis module, as illustrated in FIG. 3.

[0181] Additionally, an input table in which preset parameter values ​​are entered for parameters or a change value input table for changing preset parameter values ​​can be displayed in section 350.

[0182] The parameter values ​​for the parameters are used in signal processing of the target presence / absence reading software or serve as a basis for judgment by signal processing.

[0183] In one embodiment, the parameter may be (i) a parameter defining a criterion for determining whether the size of a data set obtained during the amplification process of a plurality of target nucleic acid molecules exceeds a threshold, or (ii) a parameter defining a criterion for determining the presence or absence of a target nucleic acid molecule using the number of cycles at which the size of a data set obtained during the amplification process of a plurality of target nucleic acid molecules reaches the threshold. For example, the parameter may be used for mathematical processing of amplification reaction result data or as a criterion for reading the presence or absence of a target nucleic acid molecule in a sample. The criterion may include a threshold intensity value of a signal used to determine the Ct (Threshold Cycle) of the sample.

[0184] In one embodiment, the parameter may be at least one parameter selected from the group consisting of:

[0185] (i) a parameter representing the relationship between the magnitude of the signal detected at a relatively high detection temperature and the magnitude of the signal detected at a relatively low detection temperature in a data set obtained during the amplification process of multiple target nucleic acid molecules,

[0186] (ii) a parameter defining a standardization coefficient for normalizing multiple data sets obtained during the amplification process of multiple target nucleic acid molecules;

[0187] (iii) a parameter defining a criterion for determining the absence of a target nucleic acid molecule using the maximum derivative of a sigmoid fitting curve applied to multiple data sets obtained during the amplification process of multiple target nucleic acid molecules;

[0188] (iv) A parameter defining a criterion for determining the presence of a target nucleic acid molecule using a parameter representing the maximum derivative of a sigmoid fitting curve applied to a plurality of data sets obtained during the amplification process of a plurality of target nucleic acid molecules,

[0189] (v) a parameter defining a criterion for determining the modification of a sigmoid fitting curve applied to multiple data sets obtained during the amplification process of multiple target nucleic acid molecules;

[0190] (vi) parameters defining the starting cycles required to perform regression analysis on multiple data sets obtained during the amplification process of multiple target nucleic acid molecules;

[0191] (vii) a parameter indicating the minimum number of final cycles required to perform regression analysis on multiple data sets obtained during the amplification process of multiple target nucleic acid molecules;

[0192] (viii) a parameter defining a criterion for determining the absence of a target nucleic acid molecule using a fitting accuracy obtained by applying a regression model applied to a portion of a plurality of data sets obtained in the process of amplifying a plurality of target nucleic acid molecules, (ix) a parameter defining a criterion for determining the absence of a target nucleic acid molecule using a fitting accuracy obtained by applying a regression model diagnosis to the entirety of a plurality of data sets obtained in the process of amplifying a plurality of target nucleic acid molecules, and

[0193] (x) A parameter defining a criterion for determining the absence of a target nucleic acid molecule by using a mathematical operation of the maximum and minimum values ​​of the sizes of multiple data sets obtained during the amplification process of multiple target nucleic acid molecules or a mathematical operation of the last cycle value and the minimum value.

[0194] These parameters can be referenced by the sub-modules described above during processing and analysis.

[0195] In the Assay component generation device (100) according to the present disclosure, a first analysis condition including a processing / analysis module, each sub-module, and a value of a parameter can be stored in the analysis information.

[0196] As described above, the values ​​of the processing / analysis module and parameters represent analysis conditions for analyzing the amplification reaction result data of the performance experiment.

[0197] Based on this, the Assay component generation device (100) can provide an interface that can generate multiple analysis conditions so that analysis can be performed by variously changing the values ​​of processing / analysis modules and parameters in the process of analyzing the amplification reaction result data for each performance experiment.

[0198] The presence / absence analysis process involves various analysis conditions that require the setting of processing / analysis modules and parameter values. However, unless you're an experienced developer, accurately setting the processing / analysis modules and parameter values ​​used in the presence / absence analysis process during the assay development process can be challenging.

[0199] Therefore, conventional real-time PCR reaction devices display presence / failure reading results based on the values ​​of reference parameters based on preset processing / analysis modules.

[0200] However, in order to find the optimal processing / analysis module and parameter values ​​for each assay, the developer needs to check the presence / absence reading results while changing all or part of the processing / analysis module and / or parameter values ​​for each assay, and check what differences exist compared to the actual confirmed presence / absence reading results. In addition, during this process, the developer also needs to recognize the relationship between the values ​​of each parameter, the changes for each parameter, and the values ​​of a specific processing / analysis module and / or a specific parameter that have a major impact on the presence / absence reading while changing the values ​​of the processing / analysis module and / or parameter to reach the actual confirmed presence / absence (positive / negative) reading results. However, since the conventional real-time PCR reaction device only displays the analyzed presence / absence reading results using the preset reference setting values, it was necessary to consider the processing / analysis module that processes and analyzes the amplification reaction result data using the actual parameter values. For this reason, the Assay component generation device (100) according to the present disclosure generates various analysis conditions to simultaneously confirm or compare the analysis results (presence / absence reading results) or changes (amplification curves) by changing the values ​​of processing / analysis modules and parameters that have a major influence on the presence / absence reading results, thereby enabling comparison and confirmation of the values ​​of processing / analysis modules and / or parameters for each analysis condition.

[0201] Accordingly, according to the present disclosure, the Assay component generation device (100) can allow any one of a plurality of processing / analysis modules to be additionally selected even after the first analysis condition is stored in the analysis information. The values ​​of parameters to be applied to each sub-module of the additionally selected processing / analysis module can be set, and a second analysis condition including the processing / analysis module, each sub-module, and the values ​​of the parameters can be stored in the analysis information. Of course, it will be apparent that the Assay component generation device (100) can generate not only the first and second analysis conditions described above, but also third and fourth analysis conditions.

[0202] In this way, the multiple analysis conditions stored in the analysis information are as shown in the list (320) of FIG. 3 in the interface provision screen (300), and each analysis condition in the list (320) of additionally generated analysis conditions can be copied, and by calling any one of the copied analysis conditions, all or part of the analysis conditions can be modified to create another analysis condition.

[0203] The Assay component generation device (100) according to the present disclosure analyzes the amplification reaction result data of a target performance experiment by allowing a user to select a processing / analysis module through an interface providing screen (300), set the values ​​of parameters referenced by sub-modules included in the selected processing / analysis module, and set the values ​​of parameters according to the reaction vessel type described later, thereby generating a plurality of analysis conditions in various ways during the analysis process and confirming the analysis results for each analysis condition. At this time, the above-described copy function can be used to generate new analysis conditions by changing some or all of the previous analysis conditions. In this case, the analysis results to which the previous analysis conditions were applied and the analysis results to which the changed analysis conditions were applied can be quickly compared and confirmed.

[0204] Rather than performing the selection of processing / analysis modules and the setting of values ​​of parameters for each reaction vessel in a standardized order through the interface provision screen (300), when using the copy function, only the desired values ​​can be selectively and quickly and easily changed based on the preset analysis conditions.

[0205] Meanwhile, it will be obvious that the nth analysis condition, in which the values ​​of the parameters driven by the sub-modules are set, can be further stored in the analysis information.

[0206] These analysis conditions can be stored in the analysis information. As illustrated in FIG. 3, multiple analysis conditions (first and second conditions) can be stored in the analysis information (310).

[0207] Analysis information (310) may include, for example, at least one analysis condition for processing and analyzing amplification reaction result data of a performance experiment.

[0208] As illustrated in FIG. 3, analysis information (310) may be, for example, a folder that contains at least one or more analysis conditions by categorizing them, and in the interface provision screen (300), a list (320) for at least one analysis condition may be displayed in the analysis information display section. In addition, analysis conditions and analysis results for at least one analysis condition selected from the list (320) may be displayed in the analysis information display section.

[0209] In this way, the Assay component creation device (100) can provide a section for displaying analysis information through an interface provision screen (300). Depending on the embodiment, the section for displaying analysis information can display analysis conditions selected from among analysis conditions displayed in a list, experimental equipment used in a performance experiment, amplification reaction result data obtained by performing an amplification reaction on a reaction plate by the experimental equipment, information on target nucleic acid molecules for each detection channel for the reaction plate, and correct answer data results of the amplification reaction result data.

[0210] Meanwhile, when a selection is made for one of the multiple processing / analysis modules through the interface provision screen (300), the value of the parameter to be applied to each sub-module of the selected processing / analysis module is set. At this time, the value of the parameter may be a value of a preset parameter for each sub-module, or a value of a parameter set by inputting a change value of a preset parameter value.

[0211] In one embodiment, when the value of a preset parameter is used as the value of a parameter of an analysis condition stored in the analysis information, the interface provision screen (300) of the Assay component creation device (100) displays the value of the preset parameter for the parameter of each sub-module.

[0212] Alternatively, if a change value for a preset parameter needs to be entered, the interface provision screen (300) of the Assay component creation device (100) can receive a change value for each parameter.

[0213] When a change value is input, analysis conditions that allow the user to identify the input change value can be created and stored in the analysis information using color and / or a shape (identifier) ​​to distinguish it from the value of a preset parameter. For example, the output color of the change value resulting from a change in a preset parameter value can be changed to allow the user to identify the change value. A shape (identifier) ​​can be additionally output on the output screen of the change value resulting from a change in a preset parameter value to allow the user to identify the change value.

[0214] On the other hand, the Assay component generation device (100) can display the analysis results obtained by processing and analyzing the amplification reaction result data using the first analysis condition, and can display the analysis results obtained by processing and analyzing the amplification reaction result data using the second analysis condition.

[0215] Of course, it will be obvious that the Assay component generation device (100) can display analysis results for not only the first and second analysis conditions described above, but also the third, fourth...n analysis conditions.

[0216] The analysis results for each analysis condition may include amplification reaction result data for the reaction plate used in the performance experiment, amplification reaction result data analyzed under the analysis conditions, raw data for the amplification reaction result data, corrected data from which a baseline is subtracted to remove a background signal value from the raw data, and the label data results indicating the presence or absence of the target for each of the plurality of target nucleic acid molecules for the performance experiment.

[0217] The display of the analysis results can be performed by calculating the analysis conditions in the processor of the Assay component generation device (100).

[0218] Alternatively, depending on the embodiment, the display of the analysis results may be performed by providing the first and / or second analysis conditions to a computation server that computes the analysis conditions, and performing computations for processing and analysis on the computation server to return the first analysis result and the second analysis result for the first and / or second analysis conditions to the device.

[0219] In addition, in the Assay component generation device (100), the analysis result for the first analysis condition can be displayed as the correct data result of the amplification reaction result data, and the analysis result for the second analysis condition can be displayed as the correct data result of the amplification reaction result data.

[0220] In order to optimize the performance of the Assay, the optimal processing / analysis module and parameter values ​​are designed in this process. In the performance optimization process of the detection reagent, the process of selecting the values ​​of the parameters referenced in the pre-designated processing / analysis module is repeatedly performed until the performance of the Assay satisfies a predetermined standard, and if the performance does not meet the standard even after repeated performance, the process of redesigning the pre-designated processing / analysis module to generate the processing / analysis module is performed. The design of the processing / analysis module and parameter values ​​to optimize the performance of the Assay in the Assay component generation device (100) depends on the results of experimental data analysis, and the experimental data analysis can be performed as follows. According to one embodiment, the experimental data analysis can be performed by comparing the prepared reaction vessel with the correct data labeled as positive and negative based on the configuration information for the panel on which the amplification reaction was performed.

[0221] The correct answer data refers to the result actually confirmed by the experimenter who adjusted the concentration of the target nucleic acid molecule for each well of the reaction vessel, and can use the result determined as actual positive or negative from an actual clinical experiment (e.g., amplification reaction), but is not limited thereto. The correct answer data can be called up through the “Label On” button in Fig. 3. For example, when analyzing experimental data, the result of analyzing the experimental data using the pre-designated processing / analysis module and the values ​​of the parameters of the processing / analysis module is compared with the correct answer data, and whether the samples labeled as positive and the samples labeled as negative in the correct answer data match the results of the experimental data analysis can be compared and analyzed. The pre-designated processing / analysis module is one whose module and parameter values, i.e., the values ​​of the parameters, are determined in advance. For example, the values ​​of the processing / analysis module and parameters for a respiratory detection reagent, the values ​​of the processing / analysis module and parameters for intestinal bacteria, or the values ​​of the processing / analysis module and parameters for sexually transmitted diseases may be each packaged, and there may be a pre-designated processing / analysis module and parameter values ​​specific to each molecular diagnostic assay.

[0222] In the present disclosure, experimental data analysis can be performed by applying amplification reaction result data obtained through an amplification reaction for a sample provided with such correct answer data to values ​​of pre-designated processing / analysis modules and parameters, and comparing the results with the correct answer data. Based on the results of the experimental data analysis, it can be determined whether to generate an assay component using the values ​​of pre-designated processing / analysis modules and parameters.

[0223] Correct answer data can be defined as a collection of correct answers for which the presence or absence of the target nucleic acid molecule in each sample is confirmed as positive or negative. If the correct answer (positive or negative) for each reaction vessel that received the sample is confirmed for each reaction vessel in the entire 96-well plate, it can be considered correct answer data for the corresponding 96-well plate. This correct answer data can be obtained for each well by labeling by the researcher who directly adjusted the concentration of the target nucleic acid in each reaction vessel.

[0224] In one embodiment, the correct answer data may be displayed as the number of positives confirmed as positive, the number of negatives confirmed as negative, and the number of grays confirmed as gray (positive or negative) in a 96 well plate.

[0225] According to one embodiment, the correct answer data may be displayed on the plate image as illustrated in 352 of FIG. For each well on the plate, the correct answer (positive or negative gray) may be displayed in a color that can distinguish the positive / negative (e.g., red for positive, blue for negative), or may be displayed as an icon or text. In this way, the correct answer data may be displayed in various ways. In addition, the correct answer data may utilize data labeled (positive or negative or gray) by the researcher who adjusted the concentration of the target nucleic acid for each reaction vessel as described above, or may utilize the actual positive or negative determination result from an actual clinical trial (e.g., amplification reaction), but is not limited thereto.

[0226] In the present disclosure, a data set obtained through an amplification reaction for a sample that provides such correct answer data is subjected to a presence / absence reading process, and the application results are compared with the correct answer data. The processing / analysis module and parameter values ​​are determined so that the data converges to the correct answer for each well, and the values ​​can be stored as analysis conditions. The analysis conditions at this time can be generated by variously changing the processing / analysis module and parameter values ​​as described above. The analysis information can be stored by classifying various analysis conditions for processing and analyzing the amplification reaction result data in a performance experiment.

[0227] For example, in the present disclosure, the analysis information may be a folder storing a plurality of analysis conditions, and each analysis condition may be a processing / analysis execution file stored in the analysis information folder.

[0228] For this reason, in the Assay component generation device (100) according to the present disclosure, various processing / analysis tasks can be performed independently for each analysis condition.

[0229] Through this, users can save analysis conditions by changing the values ​​of processing / analysis modules and / or parameters in various ways, and perform analysis according to analysis conditions, thereby checking various analysis results, thereby performing analysis tasks more easily and quickly.

[0230] At this time, each analysis condition stored in the analysis information may have different processing / analysis module and / or parameter values. This analysis information can be generated for multiple performance experiments.

[0231] In addition, each analysis condition of the saved analysis information may have different parameter values ​​if the processing / analysis module selected through the interface provision screen (300) and the additionally selected processing / analysis module are the same.

[0232] The Assay component generation device (100) according to the present disclosure can generate an Assay component based on analysis conditions selected from among analysis conditions stored in analysis information. The selected analysis conditions can be called for each performance experiment to analyze target amplification reaction result data. The selected analysis conditions can be applied to multiple performance experiments.

[0233] The Assay component may include processing / analysis modules, parameter values, and Assay information. Additionally, the Assay component may exist as a single assembly, with processing / analysis modules and parameter values ​​for each molecular diagnostic Assay.

[0234] For generating an assay component, the assay component generating device (100) according to the present disclosure provides a data input screen (500) as shown in FIG. 5.

[0235] Figure 5 shows an input screen for creating an Assay component.

[0236] Specifically, a data input screen (500) is displayed for receiving data (name, version, intended use, region, Ignore IC, PCR Instrument SW, etc.) for the Assay component.

[0237] In the data input screen (500), data (name, version, intended use, region, Ignore IC, PCR Instrument SW, etc.) for the Assay component generated from the Assay information describing multiple target nucleic acid molecules for the Assay component are entered, and at least one or more analysis conditions stored and / or selected in the Assay are displayed. Among the displayed analysis conditions, each analysis condition for each type of reaction vessel included in the reaction plate used in the performance test can be used to generate the results based on each Assay.

[0238] The Assay component may include Assay information. The Assay information may include at least one of: information describing a plurality of target nucleic acid molecules for a molecular diagnostic Assay; information matching the values ​​of sub-modules and parameters referenced by the sub-modules to each target nucleic acid molecule; or information defining the execution order of sub-modules for processing and analysis.

[0239] Information describing multiple target nucleic acid molecules for a molecular diagnostic assay may include information describing genes or analytes of the multiple target nucleic acid molecules. This assay information may be used to display the properties of assay components in target presence / absence detection software.

[0240] Meanwhile, the Assay component generation device (100) according to the present disclosure can select the reaction vessel type (340) included in the reaction plate used in the performance experiment through the interface provision screen (300). In this case, each of the first and second analysis conditions may include values ​​of different parameters for different reaction vessel types.

[0241] Additionally, the Assay component generation device (100) may further include an amplification reaction protocol for obtaining amplification reaction result data from a nucleic acid detection device (200) in the Assay component, a Run Script (which may be a code including a command for re-execution of specific sub-modules in a processing / analysis module reflecting the Assay characteristics), and Interpretation Rules.

[0242] Since the amplification reaction result data of a performance experiment differs depending on the reaction vessel type, the analysis conditions must be generated separately for each reaction vessel type. For example, in one performance experiment, the analysis conditions for the amplification reaction result data for a reaction plate including a reaction vessel using an 8-Cap Strip, the analysis conditions for the amplification reaction result data for a reaction plate including a reaction vessel using a Sealing Film, and the analysis conditions for the amplification reaction result data for a reaction plate including a reaction vessel using a tube cannot be the same.

[0243] Fig. 7 is a drawing showing a reaction vessel selection window provided in an interface provision screen (300) of an assay component generation device (100). Using the screen illustrated in Fig. 7, a reaction vessel included in a reaction plate can be selected in a corresponding performance experiment to generate analysis conditions.

[0244] The analysis conditions selected when generating an Assay component in the Assay component generation device (100) are analysis conditions of one of the analysis information applied to the amplification reaction result data of the corresponding performance experiment, and if there are multiple reaction vessel types, they should be selected for each reaction vessel type.

[0245] An Assay component can contain at least one Assay.

[0246] Each assay may include a processing / analysis module and a processing / analysis component including values ​​for the above parameters, and analysis conditions including an amplification reaction protocol for a performance experiment. Analysis conditions exist for each type of reaction vessel used in the performance experiment.

[0247] Accordingly, as illustrated in 510 of FIG. 5, the Assay component generation device (100) generates an Assay component for each molecular diagnostic Assay, for example, to generate Assay components for molecular diagnostic Assays 1 to 3, the Assay component generation device (100) can individually generate an Assay 1 component for molecular diagnostic Assay 1, an Assay 2 component for molecular diagnostic Assay 2, and an Assay 3 component for molecular diagnostic Assay 3.

[0248] The processing / analysis module for each molecular diagnostic assay component varies depending on the characteristics of the target detection reagent. This is because the order of submodules included in the processing / analysis module, as well as the addition or exclusion of certain submodules, may vary depending on the characteristics of the detection reagent. Even when the processing / analysis modules for some molecular diagnostic assays are identical, the parameter values ​​of the processing / analysis module may differ.

[0249] The processing / analysis module may be a separate module provided for each of the plurality of target nucleic acid molecules. That is, when the target nucleic acid molecules are Target 1, Target 2, and Target 3, the processing / analysis module for Target 1, the processing / analysis module for Target 2, and the processing / analysis module for Target 3 may be different from each other. The processing / analysis module may include separate sub-modules that are individually used for each of the plurality of target nucleic acid molecules.

[0250] Additionally, when an Assay component includes at least two processing / analysis modules, each of the at least two processing / analysis modules may include at least some sub-modules that are identical to each other. Since some of the sub-modules may be implemented or fine-tuned differently for each target, some may be identical or some may be different for each target.

[0251] In one embodiment, the plurality of target nucleic acid molecules may include a common module provided for some of the target nucleic acid molecules and an individual module provided for some of the target nucleic acid molecules.

[0252] Information defining the execution order of sub-modules for processing and analysis may include information regarding the order in which the sub-modules should be executed for each target nucleic acid molecule. In this case, the execution order for processing and analysis may be predefined for each of a plurality of target nucleic acid molecules. Information defining the execution order for processing and analysis may include information regarding the order in which the sub-modules should be executed for each target nucleic acid molecule. In the present disclosure, the execution order for processing and analysis may be predefined for each of a plurality of target nucleic acid molecules.

[0253] Meanwhile, the Assay component generation device (100) according to the present disclosure automatically stores processing / module and parameter values ​​for each analysis condition, and these are used for processing amplification reaction result data. The analysis conditions entered through the interface provision screen (300) can be used as a reference for performance experiment analysis required for Assay development, and thus can be matched with metadata based on the target Assay.

[0254] As described above, the display of the analysis results can be performed by providing the first and / or second analysis conditions (the third analysis condition...the nth analysis condition) to a calculation server that calculates the analysis conditions, and returning the analysis results for the analysis conditions from the calculation server to the Assay component generation device (100).

[0255] The Assay component generation device (100) according to the present disclosure can build only individual application system functions for target analysis conditions when performing analysis with analysis conditions applied without a physical server using a known lambda function service.

[0256] Additionally, the metadata described below can be used for developing a processing / analysis module in a processing / analysis component providing device connected to an assay component generation device (100).

[0257] Metadata may also be provided from the operation server described above.

[0258] The amplification reaction result data of the performance experiment is stored as metadata for the target Assay, and the metadata can be matched with analysis information including at least one analysis condition.

[0259] Analysis conditions matched with metadata can be stored in a standardized format at a specified location in the data storage structure.

[0260] In addition, the Assay component generation device (100) according to the present disclosure acquires amplification reaction result data for a target nucleic acid molecule, which is one of the results of a plurality of performance experiments, matches the acquired amplification reaction result data with metadata, and stores the amplification reaction result data matched with the metadata in a predetermined location in the data storage structure. At this time, the amplification reaction result data is also stored in a standardized format. For example, in the amplification reaction result data, the signal intensity, which is a signal value at a specific cycle, must be input in units called RFU and must be input for each cycle, and in the case of an amplification graph, it must be displayed in a color specified in advance for each channel, and the scale of the amplification graph may be normalized.

[0261] Metadata is data about data stored in a predetermined location in a pre-designed data storage structure, and according to one embodiment, various data derived during the Assay development process, for example, each data input through the interface provision screen (300), can be stored as metadata in a standardized format.

[0262] FIG. 6 is a flowchart for explaining an assay component generation method in an assay component generation device (100) according to one embodiment.

[0263] Referring to FIG. 6, a method performed in a device for generating an Assay component is performed by a computer device, and provides a plurality of processing / analysis modules based on the type of Assay at S610. The processing / analysis modules include sub-modules for processing and analyzing the amplification reaction result data of the performance experiment.

[0264] In S620, one of the multiple processing / analysis modules is selected.

[0265] The values ​​of parameters to be applied to each sub-module of the processing / analysis module selected in S630 are set.

[0266] In S640, the processing / analysis module, each of the sub-modules and the first analysis condition including the values ​​of the parameters are stored in the analysis information.

[0267] In the S650, one of the multiple processing / analysis modules is additionally selected.

[0268] The values ​​of parameters to be applied to each sub-module of the processing / analysis module additionally selected in S660 are set.

[0269] In S670, the processing / analysis module, each of the sub-modules and the second analysis condition including the values ​​of the parameters are stored in the analysis information.

[0270] In S680, the selected analysis conditions stored in the analysis information are created as an Assay component.

[0271] Since the operation of each step illustrated in Fig. 6 is identical to the function of the Assay component generation device (100) discussed above, the description of Figs. 1 to 5 described above will be used.

[0272] The present disclosure provides a memory storing at least one instruction; And a method using a memory, a processor, and one or more programs stored in the memory and configured to be executed by the processor, wherein the at least one instruction is executed by the processor, the method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development for a plurality of target nucleic acid molecule Assays, the method being performed by a computer device, and providing a plurality of processing / analysis modules based on the type of the Assay, the processing / analysis modules including sub-modules for processing and analyzing amplification reaction result data of the performance experiment, wherein one of the plurality of processing / analysis modules is selected, and a value of a parameter to be applied to each sub-module of the selected processing / analysis module is set, and a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in the analysis information, and one of the plurality of processing / analysis modules is additionally selected, and a value of a parameter to be applied to each sub-module of the additionally selected processing / analysis module is set, and a second analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in the analysis information, A computer device that performs a step of generating an analysis condition selected from among the analysis conditions stored in the above analysis information as an Assay component;

[0273] A computer-readable recording medium storing a computer program, wherein when the computer program is executed by one or more processors included in a computer device,

[0274] A method using a memory, a processor, and one or more programs stored in the memory and configured to be executed by the processor, the method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development of a plurality of target nucleic acid molecule Assays, the method comprising the steps of: providing a plurality of processing / analysis modules based on the type of the Assay; wherein the processing / analysis modules include sub-modules for processing and analyzing amplification reaction result data of the performance experiment; selecting any one of the plurality of processing / analysis modules; setting a value of a parameter to be applied to each sub-module of the selected processing / analysis module, and storing a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter in analysis information; further selecting any one of the plurality of processing / analysis modules; setting a value of a parameter to be applied to each sub-module of the further selected processing / analysis module, and storing a second analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter in the analysis information; A computer program that is performed, including a step of generating an analysis condition selected from among the analysis conditions stored in the above analysis information as an Assay component;

[0275] A computer-readable recording medium storing a computer program, wherein the computer program is executed by one or more processors included in a computer device,

[0276] A method using a memory, a processor, and one or more programs stored in the memory and configured to be executed by the processor, the method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development of a plurality of target nucleic acid molecule Assays, the method comprising the steps of: providing a plurality of processing / analysis modules based on the type of the Assay; wherein the processing / analysis modules include sub-modules for processing and analyzing amplification reaction result data of the performance experiment; selecting any one of the plurality of processing / analysis modules; setting a value of a parameter to be applied to each sub-module of the selected processing / analysis module, and storing a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter in analysis information; further selecting any one of the plurality of processing / analysis modules; setting a value of a parameter to be applied to each sub-module of the further selected processing / analysis module, and storing a second analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter in the analysis information; A computer-readable recording medium that includes a step of generating an analysis condition selected from among the analysis conditions stored in the above analysis information as an Assay component;

[0277] These programs can also be stored on a computer-readable recording medium to implement functions in a specific manner. Furthermore, terms such as "include," "comprise," or "have" described above, unless specifically stated otherwise, imply that the corresponding component may be included, and therefore should be interpreted as including other components rather than excluding other components. All terms, including technical or scientific terms, unless otherwise defined, have the same meaning as commonly understood by a person of ordinary skill in the art to which this disclosure pertains. Commonly used terms, such as terms defined in dictionaries, should be interpreted to be consistent with their contextual meaning in the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined in this disclosure.

[0278] The above description is merely an example of the technical idea of ​​the present disclosure, and those skilled in the art to which the present disclosure pertains will appreciate that various modifications and variations can be made without departing from the essential characteristics of the present disclosure. Therefore, the embodiments disclosed in the present disclosure are not intended to limit the technical idea of ​​the present disclosure, but rather to explain it, and the scope of the technical idea of ​​the present disclosure is not limited by these embodiments. The scope of protection of the present disclosure should be interpreted by the claims below, and all technical ideas within a scope equivalent thereto should be interpreted as being included in the scope of rights of the present disclosure.

[0279] While specific aspects of the present disclosure have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred implementation examples and do not limit the scope of the present disclosure. Therefore, the substantial scope of the present disclosure is defined by the appended claims and their equivalents.

Claims

1. A computer implementation method for generating an assay component used for analysis of a performance experiment required for developing an assay for detecting multiple target nucleic acid molecules, through an assay component generating device including a processor and a memory operably coupled thereto, wherein the memory stores instructions executable by the processor, and the assay component generating method includes the following steps: A step of providing a plurality of processing / analysis modules based on the type of the above Assay; the processing / analysis module includes sub-modules for processing and analyzing the amplification reaction result data of the performance experiment, A step in which one of the above processing / analysis modules is selected; A step of setting the values ​​of parameters to be applied to each sub-module of the selected processing / analysis module; A step in which a first analysis condition including the processing / analysis module, each sub-module, and the values ​​of the parameters is stored in the analysis information; A step in which one of the above processing / analysis modules is additionally selected; A step of setting the values ​​of parameters to be applied to each sub-module of the additionally selected processing / analysis module; A step in which a second analysis condition including the values ​​of the processing / analysis module, each sub-module, and the parameters is additionally stored in the analysis information; A step of selecting a final analysis condition from among the first and second analysis conditions stored in the above analysis information; and A step of generating an Assay component using the above final analysis conditions; including; 2. In the first paragraph, the value of the set parameter is, The values ​​of the preset parameters for each of the above sub-module parameters, or A method for creating an Assay component characterized in that the value of the set parameter is input by changing the value of the above-mentioned preset parameter.

3. In the second paragraph, the method, A step of displaying the values ​​of the preset parameters for the parameters of each sub-module when the values ​​of the preset parameters are used as the values ​​of the parameters of the analysis conditions stored in the analysis information; or, A method for creating an Assay component, characterized in that it further includes a step of receiving a change value when the change value must be entered.

4. In the first paragraph, the step of storing A process of displaying the analysis results obtained by processing and analyzing the amplification reaction result data using the first analysis condition; and An assay component creation method characterized by additionally including a process of processing and analyzing the amplification reaction result data using the second analysis condition and displaying the analysis result.

5. In paragraph 4, A process in which the analysis results for the first analysis condition are displayed as correct data results of the amplification reaction result data; and An Assay component creation method characterized by further including a process in which the analysis result for the second analysis condition is displayed as the correct data result of the amplification reaction result data.

6. In paragraph 5, each of the analysis results is: A method for generating an Assay component, characterized in that it includes amplification reaction result data for a reaction plate used in the above performance experiment, processed / analyzed data analyzed under the above analysis conditions, raw data for the amplification reaction result data, corrected data from which a baseline is subtracted to remove a background signal value from the raw data, and the label data result indicating the presence or absence of a target for each of the plurality of target nucleic acid molecules for the above performance experiment.

7. In the first paragraph, the method, A step of selecting the type of reaction vessel included in the reaction plate used in the above performance experiment; further comprising; A method for generating an assay component, wherein each of the first and second analysis conditions includes values ​​of different parameters for different reaction vessel types.

8. In paragraph 1, each analysis condition of the stored analysis information is: A method for creating an Assay component, characterized in that the values ​​of the parameters are different when the above-mentioned selected processing / analysis module and the above-mentioned additionally selected processing / analysis module are the same.

9. In paragraph 1, the selected analysis conditions are: A method for generating an assay component characterized in that any one of the above analysis conditions is applied to the above amplification reaction result data.

10. In the first paragraph, the method, A step of providing a section for displaying the above analysis information; the section for displaying the above analysis information displays the analysis conditions selected from the analysis conditions, the experimental equipment used in the performance experiment, the amplification reaction result data in which the experimental equipment performed an amplification reaction on the reaction plate, the information on the target nucleic acid molecule for each detection channel for the reaction plate, and the correct data result of the amplification reaction result data. A method for creating an Assay component, characterized in that the above analysis conditions and the analysis results for the above analysis conditions are displayed together within the section for displaying the analysis information.

11. In the 10th paragraph, in the section for displaying the analysis information, A method for creating an Assay component, characterized in that a list of at least one analysis condition is displayed, and analysis conditions and analysis results for at least one analysis condition selected from the list are displayed.

12. In paragraph 11, A method for creating an Assay component, characterized in that each analysis condition in the above list can be copied, and by calling any one of the copied analysis conditions, all or part of the analysis conditions are modified to create another analysis condition.

13. In paragraph 1, The above performance test includes multiple performance tests, The method further includes a step of analyzing the target amplification reaction result data by calling the selected analysis conditions for each performance experiment; A method for creating an Assay component, characterized in that the above-mentioned selected analysis conditions are applied to each of the plurality of performance experiments.

14. In paragraph 1, A step of storing the amplification reaction result data of the above performance experiment as metadata for the target Assay; and A method for creating an Assay component, characterized in that it further comprises a step of matching the above analysis information and the above metadata.

15. In the first paragraph, the step of providing the processing / analysis module comprises: A method for generating a component for an assay, characterized in that a library file of one processing / analysis module selected from a plurality of processing / analysis modules provided through a processing / analysis component providing device that generates the processing / analysis module is combined with a custom code for each assay type and received as a processing / analysis component.

16. In paragraph 5, the step of displaying the analysis results is: A method for generating an Assay component, characterized in that the first and / or second analysis conditions are provided to a calculation server that calculates the analysis conditions, and the calculation server performs calculations for the processing and analysis and returns the first analysis result and the second analysis result for the first and / or second analysis conditions to the device.

17. In the first paragraph, the step of creating the Assay component comprises: A step of providing a screen for inputting data for an Assay component generated from Assay information describing the plurality of target nucleic acid molecules for the Assay component; A step of displaying at least one analysis condition stored and / or selected in the Assay; and A method for generating an Assay component, characterized in that it comprises a step of generating an Assay component using the analysis conditions for each reaction vessel type included in the reaction plate used in the performance experiment based on the Assay among the indicated analysis conditions.

18. In the 17th paragraph, the Assay component, Contains at least one Assay, Each Assay comprises a processing / analysis component including the processing / analysis module and the values ​​of the parameters and an analysis condition including the amplification reaction protocol of the performance experiment, A method for generating an assay component, characterized in that the above analysis conditions exist for each type of reaction vessel used in the above performance experiment.

19. In the first paragraph, the processing / analysis module, A method for generating an assay component characterized in that it is a common module for at least two target nucleic acid molecules among the above plurality of target nucleic acid molecules.

20. In paragraph 1, If the above processing / analysis module includes two or more sub-modules, The components for the above Assay are: A method for creating an Assay component that further includes Assay information in which the execution order of sub-modules for processing and analysis is predefined.

21. In paragraph 20, The execution sequence for the above processing and analysis is: A method for generating an assay component, characterized in that each of the plurality of target nucleic acid molecules is predefined.

22. In the first paragraph, the processing / analysis module and the values ​​of the parameters are A method for creating an assay component characterized by being designed through a performance optimization process of the above assay.

23. In paragraph 22, The performance optimization process of the above Assay is: A step of repeating the determination of the values ​​of parameters referenced by the predetermined sub-modules until the performance of the above Assay satisfies a predetermined standard; A method for generating an Assay component, characterized in that if the performance of the Assay does not satisfy a predetermined standard after the above repetition is performed, further redesign of the predetermined sub-modules is performed.

24. In paragraph 23, In the process of redesigning the above-mentioned sub-modules, A method for creating an Assay component, characterized in that at least one of the following is performed: a process of modifying one or more sub-modules included in the above-mentioned pre-designated sub-modules, a process of adding sub-modules not included in the above-mentioned pre-designated sub-modules to the Assay component, and a process of deleting at least one of the sub-modules if the above-mentioned pre-designated sub-modules include two or more sub-modules or changing the execution order of the sub-modules.

25. In the first paragraph, the method, A method for generating an Assay component, characterized in that it further includes a step of further storing the nth analysis condition in which the value of the parameter driven by the sub-module is set in the analysis information.

26. Memory for storing at least one instruction; and Contains a processor, By executing at least one instruction by the processor, A method using a memory, a processor and one or more programs stored in said memory and configured to be executed by said processor, said method being performed in a device for generating an Assay component for performing analysis on performance experiments required for development of a plurality of target nucleic acid molecule Assays, said method comprising the steps of: providing a plurality of processing / analysis modules based on the type of the Assay, said processing / analysis modules being performed by a computer device; The processing / analysis module includes sub-modules for processing and analyzing the amplification reaction result data of the performance experiment, and one of the plurality of processing / analysis modules is selected, a value of a parameter to be applied to each sub-module of the selected processing / analysis module is set, a first analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in analysis information, and one of the plurality of processing / analysis modules is additionally selected, a value of a parameter to be applied to each sub-module of the additionally selected processing / analysis module is set, a second analysis condition including the processing / analysis module, each of the sub-modules, and the value of the parameter is stored in the analysis information, and a computer device that generates an analysis condition selected from among the analysis conditions stored in the analysis information as an Assay component.

27. A computer-readable recording medium storing a computer program, wherein when the computer program is executed by one or more processors included in a computer device, A method using a memory, a processor, and one or more programs stored in the memory and configured to be executed by the processor, the method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development of a plurality of target nucleic acid molecule Assays, the method comprising the steps of: providing a plurality of processing / analysis modules based on the type of the Assay; the processing / analysis modules including sub-modules for processing and analyzing amplification reaction result data of the performance experiment; A step in which one of the above processing / analysis modules is selected; A step in which the values ​​of parameters to be applied to each sub-module of the selected processing / analysis module are set, and a first analysis condition including the processing / analysis module, each sub-module, and the values ​​of the parameters are stored in the analysis information; A step in which one of the above processing / analysis modules is additionally selected; A step in which the values ​​of parameters to be applied to each sub-module of the additionally selected processing / analysis module are set, and a second analysis condition including the processing / analysis module, each sub-module, and the values ​​of the parameters is stored in the analysis information; A computer program that is performed, including a step of generating an analysis condition selected from among the analysis conditions stored in the above analysis information as an Assay component.

28. A computer-readable recording medium storing a computer program, wherein the computer program is executed by one or more processors included in a computer device. A method using a memory, a processor and one or more programs stored in said memory and configured to be executed by said processor, said method being performed in a device for generating an Assay component for performing an analysis on a performance experiment required for development of a plurality of target nucleic acid molecule Assays, said method being performed by a computer device, A step of providing a plurality of processing / analysis modules based on the type of the above Assay; the processing / analysis module includes sub-modules for processing and analyzing the amplification reaction result data of the performance experiment, A step in which one of the above processing / analysis modules is selected; A step in which the values ​​of parameters to be applied to each sub-module of the selected processing / analysis module are set, and a first analysis condition including the processing / analysis module, each sub-module, and the values ​​of the parameters are stored in the analysis information; A step in which one of the above processing / analysis modules is additionally selected; A step in which the values ​​of parameters to be applied to each sub-module of the additionally selected processing / analysis module are set, and a second analysis condition including the processing / analysis module, each sub-module, and the values ​​of the parameters is stored in the analysis information; and A step of generating an analysis condition selected from among the analysis conditions stored in the above analysis information as an Assay component; including; Computer readable recording medium.

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