Real-time RGB quantitative PCR system and control method thereof

Abstract

The invention discloses a real-time RGB quantitative PCR system and a control method thereof, and relates to the technical field of polymerase chain reaction detection, the PCR system comprises a PCR thermal cycle system, a light source, a camera or an RGB detector, a shading shell, a base, a computer and a display terminal; wherein the PCR thermal cycle system, the light source and the camera or the RGB detector are arranged on the base at fixed positions; the PCR thermal cycle system, the light source and the camera or the RGB detector are all arranged in the shading shell; and the camera or the RGB detector and the display terminal are respectively in communication connection with the computer. The PCR thermal cycle system, the light source and the camera or the RGB detector are arranged in the shading shell, so that the influence of natural light on RGB detection is isolated, and the detection accuracy is improved; after RGB detection, analysis and detection result display can be immediately carried out through a computer, in-situ detection and detection result display can be realized, and the efficiency is improved.

Description

Technical Field

[0001] The present application relates to the field of polymerase chain reaction detection technology, and in particular to a real-time RGB quantitative PCR system and a control method thereof. Background Art

[0002] Polymerase Chain Reaction (PCR) is an in vitro nucleic acid amplification technique characterized by high specificity, high sensitivity, simplicity, and speed. There are two common types of PCR instruments: conventional PCR instruments and real-time fluorescence quantitative PCR instruments. Conventional PCR instruments can only perform PCR amplification of the target nucleic acid. The PCR amplification products must be detected on other instruments and devices (such as gel electrophoresis instruments, fluorescence spectrophotometers, UV-visible spectrophotometers, circular dichroism spectrometers, dynamic light scattering instruments, and electrochemical workstations). PCR amplification and signal detection are performed on two separate instruments. Real-time fluorescence quantitative PCR instruments can simultaneously perform PCR amplification and in situ detection of the target nucleic acid, obtaining real-time fluorescence signals from the PCR amplification products and transmitting them to a computer analysis and processing system for real-time quantitative output.

[0003] However, the detection results of existing real-time fluorescence quantitative PCR instruments are still not accurate enough. Summary of the Invention

[0004] The main purpose of the embodiments of the present application is to provide a real-time RGB quantitative PCR system and a control method thereof to improve the accuracy of the detection results of a PCR instrument.

[0005] To achieve the above-mentioned object, one aspect of an embodiment of the present application provides a real-time RGB quantitative PCR system, wherein the PCR system comprises: a PCR thermal cycle system, a light source, a camera or an RGB detector, a light-shielding housing, a base, a computer, and a display terminal;

[0006] The PCR thermal cycle system, the light source, and the camera or RGB detector are all fixedly arranged on the base; the PCR thermal cycle system, the light source, and the camera or RGB detector are all arranged in the light-shielding housing;

[0007] The camera or RGB detector and the display terminal are respectively connected to the computer for communication;

[0008] The PCR thermal cycle system is used to perform PCR reaction;

[0009] The light source is used for lighting;

[0010] The camera or RGB detector is used to capture real-time RGB images of samples undergoing PCR reaction in the PCR thermal cycle system;

[0011] The computer is used to determine the R, G, and B parameters of the sample based on the RGB image; and is used to set the cycle time of the PCR reaction of the sample and control the temperature of the PCR reaction of the sample;

[0012] The display terminal is used to display the R, G, and B parameters.

[0013] In some embodiments, the PCR thermal cycle system includes a single chip microcomputer, a fan, a control button, an LED display screen, and a resistance reaction cell;

[0014] Wherein, the single chip microcomputer is used to set the cycle time of the sample for PCR reaction and control the temperature of the sample for PCR reaction;

[0015] The resistance reaction cell is used to place the sample and control the temperature of the heated sample;

[0016] The fan is used for cooling;

[0017] The operation button is used to start or stop the PCR reaction of the sample and adjust the control parameters of the PCR thermal cycle system;

[0018] The LED display screen is used to display the parameters of the PCR thermal cycle system.

[0019] In some embodiments, the light generated by the light source is at a 90-degree or other angle to the imaging end of the camera or RGB detector;

[0020] The sample is at a 90-degree angle or other angles to the imaging end of the camera or RGB detector.

[0021] In some embodiments, the camera or RGB detector is communicatively connected to the computer via a USB data cable or a wireless signal.

[0022] To achieve the above objectives, another aspect of the embodiments of the present application provides a control method for controlling a real-time RGB quantitative PCR system as described in the present application, the method comprising the following steps:

[0023] Acquire an RGB image obtained by capturing the sample undergoing PCR reaction in real time using a camera or an RGB detector;

[0024] Determine the R, G, B parameters of the sample according to the RGB image;

[0025] Displays the R, G, B parameters.

[0026] In some embodiments, determining the R, G, and B parameters of the sample according to the RGB image comprises the following steps:

[0027] RGB detection is performed on the area where the reagent exists in the sample in the RGB image to obtain the R, G, B values, the total RGB value, and the R / G, R / B or G / B ratio of the area where the reagent exists as the R, G, B parameters of the sample.

[0028] In some embodiments, displaying the R, G, and B parameters comprises the following steps:

[0029] Graphs showing changes in the R, G, B values, the RGB total value, and the R / G, R / B, or G / B ratios over time are drawn respectively and then displayed; wherein each graph is drawn and updated at a first set period.

[0030] In some embodiments, before displaying the R, G, B parameters, the method further includes the following steps:

[0031] Obtaining historical data of the R, G, B values, the total RGB value, and the R / G, R / B, or G / B ratios;

[0032] Sampling the historical data at a second set period to obtain a sampling value;

[0033] Compare the R, G, B values, the RGB total value, and the R / G, R / B, or G / B ratios obtained by the current detection with the sample values obtained by the current sampling;

[0034] If the difference obtained by comparison reaches the corresponding set threshold, the R, G, B value, the RGB total value or the R / G, R / B or G / B ratio obtained by the current detection is deleted.

[0035] In some embodiments, the method further comprises the following steps:

[0036] The R, G, and B parameters obtained within a set time period are stored; wherein the time period for the sample to undergo PCR reaction is within the set time period, and the duration of the set time period is longer than the duration for the sample to undergo PCR reaction.

[0037] The embodiments of the present application include at least the following beneficial effects:

[0038] The PCR system of the present application includes a PCR thermal cycler system, a light source, a camera or RGB detector, a light-shielding housing, a base, a computer, and a display terminal. The PCR thermal cycler system, the light source, and the camera or RGB detector are all fixedly positioned on the base. The PCR thermal cycler system, the light source, and the camera or RGB detector are all disposed within the light-shielding housing. The camera or RGB detector and the display terminal are each communicatively connected to the computer. By disposing the PCR thermal cycler system, the light source, and the camera or RGB detector within the light-shielding housing, the present application can isolate the influence of natural light on RGB detection, thereby improving detection accuracy. Furthermore, after RGB detection, the detection results can be immediately analyzed and displayed by the computer, enabling in-situ detection and real-time display of detection results, thereby improving efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0040] Figure 1 A flow chart of a control method provided in an embodiment of the present application;

[0041] Figure 2 This is an example structural diagram of a real-time RGB quantitative PCR system provided in an embodiment of the present application;

[0042] FIG3( a ) is an example diagram of the curves of R value, G value, and B value changing with PCR time provided in an embodiment of the present application;

[0043] FIG3( b ) is an example diagram of a curve showing a change in RGB total value versus PCR time provided in an embodiment of the present application;

[0044] FIG3( c ) is an example of a curve showing the change of the R / G, R / B or G / B ratio with PCR time provided in an embodiment of the present application;

[0045] Figure 4 This is an example diagram of the sampling results of the R, G, and B parameters provided in the embodiment of the present application. DETAILED DESCRIPTION

[0046] In order to make the purpose, technical solutions and advantages of the present application clearer, the present application is further described in detail below with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are only used to explain the present application and are not intended to limit the present application. When the following description refers to the drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the embodiments of the present application. They are merely examples of devices and methods consistent with some aspects of the embodiments of the present application as detailed in the appended claims.

[0047] It will be understood that the terms "first", "second", etc. used in this application may be used herein to describe various concepts, but unless otherwise specified, these concepts are not limited by these terms. These terms are only used to distinguish one concept from another. For example, without departing from the scope of the embodiments of the present application, the first information may also be referred to as the second information, and similarly, the second information may also be referred to as the first information. Depending on the context, the words "if" and "if" as used herein may be interpreted as "at the time of" or "when" or "in response to determining".

[0048] The terms "at least one", "plurality", "each", "any", etc. used in this application include "at least one", "two" or more, "plurality" or "each", "any" or "any one", "each" or "any one" as used herein.

[0049] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this application pertains. The terms used herein are for the purpose of describing the embodiments of this application only and are not intended to limit this application.

[0050] Before describing the embodiments of the present application in detail, some of the terms and related technologies involved in the embodiments of the present application are first explained as follows:

[0051] RGB defines the three basic colors (red, green, and blue) as 256 intensity levels, ranging from 0 to 255. The set of ordered triples (r, g, b) is called the RGB color model, defined as: RGB model = {(r, g, b) | 0 ≤ r ≤ 255, 0 ≤ g ≤ 255, 0 ≤ b ≤ 255}. This model defines a total of 256 × 256 × 256 = 16,777,216 colors, each represented by the form (r, g, b). When we talk about the RGB color model, we are referring to these 16,777,216 colors. Digital RGB color imaging allows for real-time imaging of samples. The effective integration of PCR amplification technology and RGB color imaging facilitates the development of novel real-time quantitative PCR thermal cycling systems. Currently, no quantitative PCR thermal cycling system capable of real-time RGB detection of samples exists.

[0052] Existing PCR machines can only perform PCR amplification of target nucleic acids. PCR amplification products must be detected using other instruments and devices (such as gel electrophoresis instruments, fluorescence spectrophotometers, UV-visible spectrophotometers, circular dichroism spectrometers, dynamic light scattering instruments, and electrochemical workstations). PCR amplification and signal detection are performed on two separate instruments. Currently, there are no new quantitative PCR instruments that can perform real-time imaging of samples.

[0053] Existing real-time digital RGB color imaging quantitative PCR instruments do not consider the impact of natural light sources on the actual R, G, and B values tested, nor do they limit the pixel range for detecting R, G, and B values. Detection should be limited to the area containing experimental sample reagents. A too large detection range of the sensor will inevitably affect the accuracy of RGB detection. Furthermore, they simply use a single-chip microcontroller to simply process the R, G, and B values and generate an image of their changes over time. If sudden errors occur during the experiment, they cannot be detected and processed in a timely manner. At the same time, the detected RGB data is not stored, which is not convenient for subsequent processing.

[0054] To address at least one technical problem in the prior art, the present application provides a real-time RGB quantitative PCR system and a control method thereof. The PCR system of the present application includes a PCR thermal cycler system, a light source, a camera or RGB detector, a light-shielding housing, a base, a computer, and a display terminal. The PCR thermal cycler system, the light source, and the camera or RGB detector are all fixedly positioned on the base. The PCR thermal cycler system, the light source, and the camera or RGB detector are all disposed within the light-shielding housing. The camera or RGB detector and the display terminal are respectively connected to the computer for communication. By disposing the PCR thermal cycler system, the light source, and the camera or RGB detector within the light-shielding housing, the present application can isolate the influence of natural light on RGB detection, thereby improving detection accuracy. Furthermore, after RGB detection, the detection results can be immediately analyzed and displayed by the computer, enabling in-situ detection and real-time display of detection results, thereby improving efficiency.

[0055] The embodiment of the present application provides a control method, which relates to the field of polymerase chain reaction detection technology. The control method provided in the embodiment of the present application can be applied to a terminal, can also be applied to a server, and can also be software running in a terminal or a server. In some embodiments, the terminal can be a smart phone, a tablet computer, a laptop computer, a desktop computer, a smart speaker, a smart watch, and a car terminal, etc., but is not limited to this; the server side can be configured as an independent physical server, or can be configured as a server cluster or distributed system composed of multiple physical servers, and can also be configured as a cloud server that provides basic cloud computing services such as cloud services, cloud databases, cloud computing, cloud functions, cloud storage, network services, cloud communications, middleware services, domain name services, security services, CDN, and big data and artificial intelligence platforms. The server can also be a node server in a blockchain network; the software can be an application that implements the control method, etc., but is not limited to the above forms.

[0056] The present application can be used in many general or special computer system environments or configurations. For example: personal computers, server computers, handheld or portable devices, tablet devices, multiprocessor systems, microprocessor-based systems, set-top boxes, programmable consumer electronics, network PCs, minicomputers, mainframe computers, distributed computing environments including any of the above systems or devices, and the like. The present application can be described in the general context of computer-executable instructions executed by a computer, such as program modules. Generally, program modules include routines, programs, objects, components, data structures, and the like that perform specific tasks or implement specific abstract data types. The present application can also be practiced in distributed computing environments in which tasks are performed by remote processing devices connected via a communication network. In a distributed computing environment, program modules can be located in local and remote computer storage media, including storage devices.

[0057] The embodiment of the present application provides a real-time RGB quantitative PCR system, wherein the PCR system comprises: a PCR thermal cycle system, a light source, a camera or an RGB detector, a light-shielding housing, a base, a computer, and a display terminal;

[0058] The PCR thermal cycle system, the light source, and the camera or RGB detector are all fixedly arranged on the base; the PCR thermal cycle system, the light source, and the camera or RGB detector are all arranged in the light-shielding housing;

[0059] The camera or RGB detector and the display terminal are respectively connected to the computer for communication;

[0060] The PCR thermal cycle system is used to perform PCR reaction;

[0061] The light source is used for lighting;

[0062] The camera or RGB detector is used to capture real-time RGB images of samples undergoing PCR reaction in the PCR thermal cycle system;

[0063] The computer is used to determine the R, G, and B parameters of the sample based on the RGB image; and is used to set the cycle time of the PCR reaction of the sample and control the temperature of the PCR reaction of the sample;

[0064] The display terminal is used to display the R, G, and B parameters.

[0065] As an optional implementation, the light source of the embodiment of the present application may include an LED lamp, a xenon lamp, or any other optional light source.

[0066] As an optional embodiment, the PCR thermal cycle system includes a single chip microcomputer, a fan, a control button, an LED display screen and a resistance reaction cell;

[0067] Wherein, the single chip microcomputer is used to set the cycle time of the sample for PCR reaction and control the temperature of the sample for PCR reaction;

[0068] The resistance reaction cell is used to place the sample and control the temperature of the heated sample;

[0069] The fan is used for cooling;

[0070] The operation button is used to start or stop the PCR reaction of the sample and adjust the control parameters of the PCR thermal cycle system;

[0071] The LED display screen is used to display the parameters of the PCR thermal cycle system.

[0072] As an optional embodiment, the light generated by the light source is at 90 degrees or other angles to the camera end of the camera or RGB detector;

[0073] The sample is at a 90-degree angle or other angles to the imaging end of the camera or RGB detector.

[0074] As an optional embodiment, the light source includes an LED lamp whose brightness can be adjusted remotely.

[0075] As an optional implementation, the camera or RGB detector is connected to the computer via a USB data cable or a wireless signal.

[0076] Reference Figure 1 The present application also provides a control method for controlling a real-time RGB quantitative PCR system as described in the present application, the method comprising the following steps S100 to S120:

[0077] S100: Acquire an RGB image obtained by real-time shooting of the sample undergoing PCR reaction by a camera or an RGB detector.

[0078] S110: Determine R, G, B parameters of the sample according to the RGB image.

[0079] Furthermore, S110 may include the following steps S111:

[0080] S111: Perform RGB detection on the area where the reagent exists in the sample in the RGB image to obtain the R, G, B values, the total RGB value, and the R / G, R / B or G / B ratio of the area where the reagent exists as the R, G, B parameters of the sample.

[0081] S120: Display the R, G, B parameters.

[0082] Furthermore, S120 may include the following steps S121:

[0083] S121: respectively plotting graphs showing changes in the R, G, B values, the total RGB value, and the R / G, R / B, or G / B ratios over time, and then displaying each of the graphs; wherein each of the graphs is plotted and updated at a first set period.

[0084] In some embodiments, before displaying the R, G, and B parameters in S120, the method of the embodiment of the present application may further include a step of removing outliers. The step of removing outliers may include the following steps:

[0085] Obtaining historical data of the R, G, B values, the total RGB value, and the R / G, R / B, or G / B ratios;

[0086] Sampling the historical data at a second set period to obtain a sampling value;

[0087] Compare the R, G, B values, the RGB total value, and the R / G, R / B, or G / B ratios obtained by the current detection with the sample values obtained by the current sampling;

[0088] If the difference obtained by comparison reaches the corresponding set threshold, the R, G, B value, the RGB total value or the R / G, R / B or G / B ratio obtained by the current detection is deleted.

[0089] Furthermore, the method of the embodiment of the present application may further include the following steps:

[0090] The R, G, and B parameters obtained within a set time period are stored; wherein the time period for the sample to undergo PCR reaction is within the set time period, and the duration of the set time period is longer than the duration of the sample to undergo PCR reaction. The solution of the embodiment of this application will be described in detail and explained in conjunction with specific application examples.

[0091] Next, the solution of the embodiment of the present application is described in conjunction with more specific implementation methods.

[0092] To address some of the challenges of the prior art, this embodiment provides a real-time RGB quantitative PCR system that allows real-time observation of the growth of PCR amplification products in sample tubes, as reflected by real-time changes in R, G, and B values. This system eliminates the need for traditional gel electrophoresis followed by detection using instruments such as a UV analyzer. This embodiment utilizes a portable PCR instrument, coupled with the provided RGB real-time detection software, to conduct PCR experiments in a darkroom constructed from a 3D-printed, light-shielding housing. This results in a more convenient and accurate real-time RGB quantitative PCR device, improving experimental throughput and operational efficiency, while offering a high degree of automation and more reliable results.

[0093] Specifically, the control method of the embodiment of the present application can be implemented through a software solution.

[0094] The software package includes:

[0095] 1. Virtual Machine: The software runs on a Linux system, while the PC environment is a Windows system. Therefore, you need to build a virtual machine on the PC to configure the Linux system for the software. First, you need to install a virtualization software to create and manage virtual machines. Here, you choose VirtualBox. Then, download the ISO image file of the Linux system from the official website. Then, create a new virtual machine in the VirtualBox software, set the memory, CPU, hard disk and other resources of the virtual machine, and then start the virtual machine. After starting the virtual machine, the system will boot from the ISO image and enter the Linux installation interface. After the installation is completed according to the instructions, you can log in through the virtual machine interface and start using the Linux operating system.

[0096] 2. User Interface (UI): The software's UI is developed in Python using Kivy, an open-source Python library for developing cross-platform graphical user interfaces (GUIs). The UI begins with a homepage with a background image and a menu. Clicking "camerax" in the menu activates the external camera or RGB detector and enters the camera or RGB detector's monitoring interface. The upper left corner displays the real-time detected R, G, and B values, the total RGB value, and the R / G, R / B, or G / B ratios. On the right side of the interface are two buttons for exporting to Excel and drawing charts.

[0097] 3. Real-time RGB Detection Software: This RGB detection software, developed in Python and based on Kivy, processes image data captured by a camera. It first captures a live image from an external camera or RGB detector and analyzes the R, G, and B values in the central window area. The software updates the R, G, B values, the total RGB value, and the R / G, R / B, or G / B ratios in real time, displaying these values in the upper left corner of the screen. The software has two function buttons on the right. The upper right button first acquires a historical record of the R, G, B values, the total RGB value, and the R / G, R / B, or G / B ratios. It then samples these values every 0.2 seconds (an optional second set period). If a sampled value differs significantly from its neighboring values, it is considered an error and discarded. The remaining values are then exported to an Excel file. A pop-up window will indicate "Excel file exported successfully" if the export is successful, while a pop-up window will indicate the error reason if the export fails. The lower right button plots the collected RGB data into a chart. It generates three types of graphs: a graph showing the R, G, and B values over time, which displays the trends of each value over time; a graph showing the trends of the RGB total value over time; and a graph showing the RGB color ratios over time, which displays the changes in the R / G, R / B, and G / B ratios over time. To avoid storing excessive historical data, and because a typical PCR experiment typically takes between one and one and a half hours, this embodiment limits the RGB recording duration to a maximum of two hours (an optional method for setting the time period). The graphs are also sampled every 1 second (an optional method for the first set period).

[0098] Next, an example of a real-time RGB quantitative PCR system provided in this embodiment is described. Specifically, the PCR system of this embodiment includes:

[0099] 1. Portable PCR instrument (PCR equipment):

[0100] This portable PCR instrument can be powered by a simple USB power adapter. Despite its small size, it is a fully functional thermal cycler, offering the functions of a conventional PCR instrument (PCR cycle times can be individually set), including ramping, cooling, and constant temperature control. It offers precise temperature control (ranging from ambient to 99°C) and accommodates five sample pools for experiments. The instrument primarily consists of a small microcontroller, a small fan, a control button, an LED display, and a resistor reaction cell. The microcontroller sets the cycle time, while a PID (Proportional-Integral-Derivative Controller) (a feedback control mechanism commonly used in industrial control systems, designed to keep the system output close to a target value. It adjusts the controlled variable to correct system errors, achieving precise control) temperature control is primarily responsible. The fan provides cooling, while the resistor reaction cell controls heating and serves as a sample holder. The control button controls the start and end of the experiment and adjusts various numerical values. The LED display displays experimental data.

[0101] 2. PC: A laptop computer (including a display terminal), mainly used to carry the RGB detection software provided in this embodiment, detect the R, G, and B values of the PCR reaction in real time, and also used to store experimental data and draw a chart showing the changes of R, G, and B values over time.

[0102] 3. Camera: The external camera is connected to the PC via USB. At the same time, the camera end is aimed at the sample part of the PCR instrument to monitor the changes of the sample during the PCR reaction in real time, and the real-time monitoring image is transmitted to the PC for processing.

[0103] 4. 3D-printed light-shielding housing and base: The 3D-printed light-shielding housing is printed with white resin and painted black to enhance its ability to block natural light. The light-shielding housing is mainly used to create a darkroom. In addition, this embodiment also provides a 3D-printed base, which is used to fix the position of the portable PCR instrument, LED light, and camera to ensure that the position is the same for each experiment, greatly reducing the impact of natural light on the accuracy of RGB detection and improving the accuracy of the experiment.

[0104] 5. LED lamp (light source): A remotely controlled LED lamp is used as a light source in a dark room. Preferably, the light source, PCR sample, and camera are at 90 degrees or other angles so that the light from the light source cannot directly enter the camera and affect the experimental accuracy.

[0105] like Figure 2As shown, a fixed base secures the LED lamp, camera, and portable PCR instrument within the darkroom created by the housing. The LED lamp and camera are positioned at a 0° angle, preventing direct light from the LED into the camera. The camera receives the light signal from the LED shining on the sample, representing the sample's color change. This change is then transmitted to a PC processor for processing. The resulting image—a real-time sample pool image and RGB detection data—is displayed on a display screen. The R, G, and B values can be graphed over time on the PC or exported to an Excel spreadsheet for further data processing.

[0106] For example, Figure 3(a) is a graph showing the changes in R value, G value, and B value with PCR time, Figure 3(b) is a graph showing the changes in the total RGB value with PCR time, and Figure 3(c) is a graph showing the changes in the R / G, R / B, or G / B ratio (R / G, R / B, or G / B) with PCR time.

[0107] like Figure 4 As shown, the Excel table saves the experimental R value, G value, B value, RGB total value, RGB average value or R / G, R / B or G / B ratio (R / G, R / B or G / B), and the data is sampled every 0.2s to prevent data from being too redundant.

[0108] The beneficial effects of this embodiment include:

[0109] A self-developed and 3D-printed light-shielding housing was used to create a darkroom, preventing RGB detection from being affected by natural light. Furthermore, a base was designed and 3D-printed to secure the portable PCR instrument, LED light, and camera, ensuring consistent positioning for each experiment and improving experimental accuracy. An RGB detection program was also provided, limiting the pixel range for RGB detection to only the area containing experimental sample reagents, improving RGB detection accuracy. This detection program not only displays the monitoring screen in real time, but also displays real-time R, G, and B values, RGB total values, and R / G, R / B, or G / B ratios, making it more intuitive and convenient. Two buttons were also provided: one for displaying a graph of RGB values changing over time, and the other for exporting the detected R, G, and B values, R / G, R / B, or G / B ratios, and RGB total values into an Excel spreadsheet, facilitating subsequent data processing. Through the above scheme, this embodiment achieves more accurate and convenient real-time RGB detection for PCR experiments.

[0110] The embodiments described in the embodiments of this application are intended to more clearly illustrate the technical solutions of the embodiments of this application and do not constitute a limitation on the technical solutions provided by the embodiments of this application. Those skilled in the art will appreciate that with the evolution of technology and the emergence of new application scenarios, the technical solutions provided in the embodiments of this application are also applicable to similar technical problems.

[0111] Those skilled in the art will understand that the technical solutions shown in the figures do not constitute a limitation on the embodiments of the present application, and may include more or fewer steps than shown in the figures, or a combination of certain steps, or different steps.

[0112] Those skilled in the art will appreciate that all or some of the steps in the methods, systems, and functional modules / units in the devices disclosed above may be implemented as software, firmware, hardware, or appropriate combinations thereof.

[0113] The terms "first", "second", "third", "fourth", etc. (if any) in the specification of the present application and the above-mentioned drawings are used to distinguish similar objects and are not necessarily used to describe a specific order or sequential order. It should be understood that the data used in this way can be interchangeable where appropriate, so that the embodiments of the present application described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions, for example, a process, method, system, product or device that includes a series of steps or units is not necessarily limited to those steps or units clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products or devices.

[0114] It should be understood that in this application, "at least one (item)" means one or more, and "plurality" means two or more. "And / or" is used to describe the association relationship of associated objects, indicating that three relationships may exist. For example, "A and / or B" can mean: only A exists, only B exists, and A and B exist at the same time, where A and B can be singular or plural. The character " / " generally indicates that the previous and next associated objects are in an "or" relationship. "At least one of the following items" or similar expressions refers to any combination of these items, including any combination of single items or plural items. For example, at least one of a, b or c can mean: a, b, c, "a and b", "a and c", "b and c", or "a and b and c", where a, b, c can be single or multiple.

[0115] The preferred embodiments of the present invention are described above with reference to the accompanying drawings, but are not intended to limit the scope of the present invention. Any modifications, equivalent substitutions, and improvements made by those skilled in the art without departing from the scope and essence of the present invention should be within the scope of the present invention.

Claims

1. A real-time RGB quantitative PCR system, characterized in that: The PCR system includes: a PCR thermal cycle system, a light source, a camera or an RGB detector, a light-shielding housing, a base, a computer, and a display terminal; The PCR thermal cycle system, the light source, and the camera or RGB detector are all fixedly arranged on the base; the PCR thermal cycle system, the light source, and the camera or RGB detector are all arranged in the light-shielding housing; The camera or RGB detector and the display terminal are respectively connected to the computer for communication; The PCR thermal cycle system is used to perform PCR reaction; The light source is used for lighting; The camera or RGB detector is used to capture real-time images of samples undergoing PCR reaction in the PCR thermal cycle system or to extract RGB signals; The computer is used to determine the R, G, and B parameters of the sample based on the RGB image; and is used to set the cycle time of the PCR reaction of the sample and control the temperature of the PCR reaction of the sample; The display terminal is used to display the R, G, and B parameters.

2. A real-time RGB quantitative PCR system according to claim 1, characterized in that: The PCR thermal cycle system includes a single chip microcomputer, a fan, a control button, an LED display screen and a resistance reaction cell; Wherein, the single chip microcomputer is used to set the cycle time of the sample for PCR reaction and control the temperature of the sample for PCR reaction; The resistance reaction cell is used to place the sample and control the temperature of the heated sample; The fan is used for cooling; The operation button is used to start or stop the PCR reaction of the sample and adjust the control parameters of the PCR thermal cycle system; The LED display screen is used to display the parameters of the PCR thermal cycle system.

3. A real-time RGB quantitative PCR system according to claim 1, characterized in that: The light generated by the light source is at 90 degrees or other angles to the camera end of the camera or RGB detector; The sample is at a 90-degree angle or other angles to the imaging end of the camera or RGB detector.

4. A real-time RGB quantitative PCR system according to claim 1, characterized in that: The camera or RGB detector is connected to the computer via a USB data line or a wireless signal.

5. A control method, characterized in that: Used to control a real-time RGB quantitative PCR system according to any one of claims 1 to 4, the method comprising the following steps: Acquire an RGB image obtained by capturing the sample undergoing PCR reaction in real time using a camera or an RGB detector; Determine the R, G, B parameters of the sample according to the RGB image; Displays the R, G, B parameters.

6. A control method according to claim 5, characterized in that: Determining the R, G, B parameters of the sample according to the RGB image comprises the following steps: RGB detection is performed on the area where the reagent exists in the sample in the RGB image to obtain the R, G, B values, the total RGB value, and the R / G, R / B or G / B ratio of the area where the reagent exists as the R, G, B parameters of the sample.

7. A control method according to claim 6, characterized in that: The display of the R, G, B parameters comprises the following steps: Graphs showing changes in the R, G, B values, the RGB total value, and the R / G, R / B, or G / B ratios over time are drawn respectively and then displayed; wherein each graph is drawn and updated at a first set period.

8. A control method according to claim 6, characterized in that: Before displaying the R, G, B parameters, the method further includes the following steps: Obtaining historical data of the R, G, B values, the total RGB value, and the R / G, R / B, or G / B ratios; Sampling the historical data at a second set period to obtain a sampling value; Compare the R, G, B values, the RGB total value, and the R / G, R / B, or G / B ratios obtained by the current detection with the sample values obtained by the current sampling; If the difference obtained by comparison reaches the corresponding set threshold, the R, G, B value, the RGB total value or the R / G, R / B or G / B ratio obtained by the current detection is deleted.

9. A control method according to any one of claims 5 to 8, characterized in that: The method further comprises the following steps: The R, G, and B parameters obtained within a set time period are stored; wherein the time period for the sample to undergo PCR reaction is within the set time period, and the duration of the set time period is longer than the duration for the sample to undergo PCR reaction.

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