PCR device calibration method, apparatus, system, PCR device, and storage medium

Abstract

Provided are a PCR device calibration method, an apparatus, a system, and a PCR device. The calibration method comprises: controlling each channel to be tested to test a calibration fluorescent material in a well to be tested, so as to obtain first fluorescence data of said well; acquiring standard fluorescence data of the calibration fluorescent material; determining the proportion between the first fluorescence data and the standard fluorescence data; according to the proportion, determining an adjustment mode for a PCR device to be calibrated, the adjustment mode being a current adjustment mode and / or a gain adjustment mode; and according to the adjustment mode, adjusting the working current and / or working gain of the PCR device to be calibrated, so as to determine a target working current and a target working gain for the PCR device to be calibrated, such that each channel to be tested performs a fluorescence test under the target working current and the target working gain. For a multi-channel PCR device, the present invention adjusts working parameters by means of different modes, thereby improving the test accuracy of the PCR device.

Description

Calibration method, device, system, PCR device and storage medium for PCR equipment

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of Chinese patent application 202311871550.6 filed on December 29, 2023, the contents of which are incorporated herein by reference. Technical Field

[0003] The present application relates to the field of fluorescence detection, and specifically to a calibration method, device, system, PCR device and storage medium for PCR equipment. Background Art

[0004] The real-time fluorescence quantitative PCR instrument (POCT) is used to detect fluorescence during the cycling process and collect fluorescence data through a computer connected to the real-time device. The data is displayed in the form of a standard curve using the developed real-time automatic analysis software. The real-time fluorescence quantitative PCR instrument (POCT) mainly uses the external standard curve method to quantitatively analyze specific DNA sequences in the test sample. At present, the real-time fluorescence quantitative PCR instrument (POCT) is widely used in many fields such as gene expression research, transgenic research, gene polymorphism research, drug efficacy assessment, pathogen detection, etc. The temperature field module and optical system of the real-time fluorescence quantitative PCR instrument (POCT) need to be monitored to achieve calibration of the instrument parameters before the gene amplification reaction, as well as real-time monitoring during the reaction process. As can be seen from the above, the PCR instrument (POCT) mainly analyzes the fluorescence data during the collection process. If the instrument performance is unqualified, it will lead to inaccurate measurements, which will have a negative impact on scientific research.

[0005] Summary of the Invention

[0006] According to various embodiments of the present application, a first aspect of the present application provides a calibration method for a PCR device, wherein the PCR device to be calibrated includes a well to be measured and at least one channel to be measured, and the calibration method includes:

[0007] Controlling each channel to be measured to detect the calibration fluorescent material of the well position to be measured to obtain first fluorescent data of the well position to be measured;

[0008] Acquiring standard fluorescence data of the calibration fluorescent product;

[0009] determining a ratio between the first fluorescence data and the standard fluorescence data;

[0010] determining an adjustment mode of the PCR device to be calibrated according to the ratio, wherein the adjustment mode is a current adjustment mode and / or a gain adjustment mode;

[0011] The operating current and / or operating gain of the PCR device to be calibrated is adjusted according to the adjustment mode to determine the target operating current and target operating gain corresponding to each channel to be tested, so that each channel to be tested performs fluorescence detection with the target operating current and the target operating gain.

[0012] A second aspect of the present application provides a calibration device for a PCR device, comprising:

[0013] a memory configured to store instructions;

[0014] The processor is configured to call instructions from a memory and implement the calibration method for a PCR device according to the first aspect when executing the instructions.

[0015] A third aspect of the present application provides a PCR device, comprising:

[0016] Hole position to be measured;

[0017] At least one channel to be tested;

[0018] And a calibration device for a PCR device according to the second aspect.

[0019] A fourth aspect of the present application provides a calibration system for a PCR device, comprising:

[0020] A PCR device to be calibrated, comprising a well to be tested and at least one channel to be tested;

[0021] And a calibration device for a PCR device according to the second aspect.

[0022] The above-mentioned scheme provided by the present application determines the current adjustment mode or gain adjustment mode by detecting the target well positions selected from the well positions to be tested, and obtaining the ratio between the actual value and the measured value of the calibration fluorescent product, thereby adjusting the operating current and / or operating gain of the PCR device to be calibrated, and each channel to be tested performs fluorescence detection based on the adjusted target operating current and target operating gain. The present application can adjust the operating parameters of single-throughput and multi-throughput multi-channel PCR devices in different modes to make the adjusted PCR devices meet the use standards and meet the fluorescence performance requirements of the instrument when it is produced, thereby improving the accuracy of PCR device detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] FIG1 schematically shows a flow chart of a calibration method for a PCR device according to an embodiment of the present application.

[0024] FIG2 schematically shows a circuit diagram of current regulation and gain regulation according to an embodiment of the present application.

[0025] FIG3 schematically shows a flow chart of a calibration method for a PCR device according to a specific embodiment of the present application.

[0026] FIG4 schematically shows a structural block diagram of a PCR device according to an embodiment of the present application.

[0027] FIG5 schematically shows a structural block diagram of a calibration device for PCR equipment according to an embodiment of the present application.

[0028] FIG6 schematically shows a schematic diagram of a calibration system for a PCR device according to an embodiment of the present application.

[0029] FIG7 schematically shows a structural diagram of a computer device according to an embodiment of the present application. DETAILED DESCRIPTION

[0030] To make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. It should be understood that the specific implementation methods described herein are only used to illustrate and explain the embodiments of the present application and are not used to limit the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0031] It should be noted that if the embodiments of the present application involve directional indications (such as up, down, left, right, front, back, etc.), the directional indications are only used to explain the relative position relationship, movement status, etc. between the various components under a certain specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indications will also change accordingly.

[0032] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present application, the descriptions of "first", "second", etc. are only for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of such features. In addition, the technical solutions between the various embodiments can be combined with each other, but they must be based on the fact that they can be implemented by ordinary technicians in this field. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by this application.

[0033] Figure 1 schematically shows a flow chart of a calibration method for a PCR device according to an embodiment of the present application. As shown in Figure 1 , an embodiment of the present application provides a calibration method for a PCR device, which may include the following steps.

[0034] A PCR device to be calibrated includes wells to be tested and at least one channel to be tested. It is understood that a PCR device, also known as a PCR amplifier, is an instrument that uses PCR technology to amplify specific DNA. A PCR device to be calibrated refers to a PCR device whose fluorescence acquisition values ​​have not been calibrated to meet the operating standards. A PCR device to be calibrated may include at least one well to be tested and at least one channel to be tested. As can be understood, a well refers to a detection unit on a PCR device for fluorescent samples, typically referring to the wells on a PCR device used to hold fluorescent samples and reaction solutions. Each well can perform an independent PCR reaction, and a multi-well design enables simultaneous testing of multiple samples. Different wells can have different throughputs. Fluorescence throughput refers to the number of samples that can be tested in a single PCR reaction. The higher the throughput, the more samples can be tested in a single PCR reaction. A channel refers to a detection channel on a PCR device for fluorescent samples, typically referring to the channel used to detect fluorescence signals. Fluorescent samples from different wells can be detected in the same channel. Wells to be tested refer to wells to be calibrated, and channels to be tested refer to the channels to be calibrated.

[0035] During the calibration of a PCR instrument, one can first select any one of the multiple test sites as the target site. Specifically, if the PCR instrument to be calibrated is a single-site thermal cycler, the single test site on the instrument is the target site. If the PCR instrument to be calibrated is a multi-site thermal cycler, any one of the test sites can be selected as the target site.

[0036] S102 , controlling each channel to be measured to detect the calibration fluorescent material of the well position to be measured to obtain first fluorescence data of the well position to be measured.

[0037] S104, obtaining standard fluorescence data of the calibration fluorescent product.

[0038] It is understood that a calibration fluorescent product, i.e., a fluorescence calibration product, generally refers to a standard sample with known fluorescence properties, which can be used to calibrate the calibration fluorescent products of a fluorescence detection instrument or method. The first fluorescence data refers to the fluorescence property data of the calibration fluorescent product tested at different wavelengths, including but not limited to fluorescence intensity, emissivity, etc. The standard fluorescence data refers to the known standard fluorescence data of the calibration fluorescent product. Specifically, the processor can control the channel to be tested to detect the calibration fluorescent product of the well to be tested, and obtain the first fluorescence data of the calibration fluorescent product of the target well to be tested under the detection of the channel to be tested.

[0039] S106 , determining a ratio between the first fluorescence data and the standard fluorescence data.

[0040] Specifically, during the fluorescence calibration process, the ratio between the first fluorescence data and the standard fluorescence data can be determined, thereby comparing the first fluorescence data of the measured calibration fluorescence product with the standard fluorescence data to determine whether the test performance of the instrument is accurate.

[0041] S108 , determining an adjustment mode of the PCR device to be calibrated according to the ratio, where the adjustment mode is a current adjustment mode and / or a gain adjustment mode.

[0042] It will be understood that the current regulation mode controls the reaction temperature and reaction rate of the PCR reaction by adjusting the current of the PCR device to be calibrated. During the PCR reaction, different temperatures affect processes such as DNA denaturation, annealing, and extension. Adjusting the current can affect the efficiency and quality of DNA amplification, thereby affecting the measured fluorescence data. The gain regulation mode controls the sensitivity and noise level of the detection system by adjusting the operational amplifier gain of the PCR device to be calibrated, thereby affecting the measured fluorescence data. Different fluorescent markers can be detected in different channels. The ratio between the actual fluorescence data and the test data can reflect the error of the PCR device to be calibrated. Therefore, for each channel to be measured, the processor can determine whether the PCR device to be calibrated should be regulated using the current regulation mode, the gain regulation mode, or both the current and gain regulation modes based on the ratio between the first fluorescence data and the standard fluorescence data. Specifically, referring to Figure 2, the current regulation mode adjusts the current of the LED lamp at the transmitting end of the PCR device to adjust the LED's luminous intensity. The gain regulation mode adjusts the signal at the receiving end of the PCR device to perform gain adjustment.

[0043] In an embodiment of the present application, determining the adjustment mode of the PCR device to be calibrated based on the ratio includes: when the ratio is within a preset ratio range, determining the adjustment mode to be a current adjustment mode; when the ratio is not within the preset ratio range and the working gain is continuously adjusted, determining the adjustment mode to be a gain adjustment mode.

[0044] In an embodiment of the present application, determining the adjustment mode of the PCR device to be calibrated according to the ratio also includes: when the ratio is not within the preset ratio range and the working gain is not continuously adjusted, determining the adjustment mode as the current adjustment mode and the gain adjustment mode.

[0045] It can be understood that the operating current refers to the current used by the PCR device to be calibrated to detect the calibration fluorescent product. The operating gain refers to the operational amplifier gain used by the PCR device to be calibrated to detect the calibration fluorescent product. When the fluorescent products of different wells to be measured are detected through the channels to be measured at the same time, the operating current and operating gain of the wells to be measured are the same. The preset ratio range refers to the ratio range set in advance by technicians based on technical experience. For example, when the ratio is [1 / 2, 2], the current regulation mode is adopted to adjust the current of the LED lamp at the transmitting end. When the ratio is not [1 / 2, 2], the gain regulation mode is adopted to adjust the gain of the receiving end signal, or the current and gain are adjusted simultaneously.

[0046] If the ratio is not within the preset range, and the operating gain can be adjusted continuously, the gain can be adjusted for coarse adjustment until the ratio is within the preset range. If the operating gain cannot be adjusted continuously, for example, each adjustment is incrementally increased or decreased, then it is possible that the previous adjustment ratio was less than 1 / 2, and the next adjustment ratio is greater than 2, making accurate adjustment impossible. In this case, the current adjustment mode can be used to adjust the current until the ratio is within the preset range. If the ratio is not within the preset range, it can be adjusted using either the gain adjustment mode or the current adjustment mode.

[0047] In an embodiment of the present application, for each channel to be tested, determining the adjustment mode of the PCR device to be calibrated according to the ratio includes: for each channel to be tested, obtaining the adjustment ratio for multiple adjustments of the working current and / or working gain; for each channel to be tested, after each adjustment of the working current and / or working gain according to the adjustment mode and the adjustment ratio, redetermining the ratio between the first fluorescence data and the standard fluorescence data detected after each adjustment; for each channel to be tested, when the corresponding ratio after each adjustment is within a preset ratio range, determining that the adjustment mode corresponding to each adjustment is the current adjustment mode; for each channel to be tested, when the corresponding ratio after each adjustment is within the preset ratio range, determining that the adjustment mode corresponding to each adjustment is the gain adjustment mode, or the gain adjustment mode and the current adjustment mode.

[0048] Specifically, for each channel to be measured, the processor can adjust the operating current and operating gain multiple times according to an adjustment ratio. The adjustment ratio can be a linear or nonlinear ratio. In the current adjustment mode, the operating current is adjusted according to the adjustment ratio. Specifically, the current operating current can be obtained by increasing or decreasing the set adjustment ratio based on the previous operating current. After each adjustment of the operating current, the first fluorescence data of the calibration fluorescent material is re-detected according to the adjusted operating current, thereby re-determining the ratio between the first fluorescence data and the standard fluorescence data. In the gain adjustment mode, the operating gain is adjusted according to the adjustment ratio, and the ratio between the first fluorescence data and the standard fluorescence data can be re-determined according to the above-mentioned method. Specifically, the current operating gain can be obtained by increasing or decreasing the set adjustment ratio based on the previous operating gain. The adjustment ratio of the operating current can be a first adjustment ratio, and the adjustment ratio of the operating gain can be a second adjustment ratio. Furthermore, if the corresponding ratio after each adjustment is within a preset ratio range, the processor can determine that the corresponding adjustment mode after the current adjustment is the current adjustment mode, i.e., the next adjustment mode is the current adjustment mode. If the corresponding ratio after each adjustment is within the preset ratio range, the processor can determine that the next adjustment mode is the gain adjustment mode. The preset ratio range is the ratio range set by technicians based on their technical experience. For example, if the corresponding ratio after adjustment is [1 / 2, 2], linear adjustment is used to adjust the current. If the corresponding ratio after adjustment is not [1 / 2, 2], linear adjustment is used to adjust the gain.

[0049] S110, adjusting the operating current and / or operating gain of the PCR device to be calibrated according to the adjustment mode to determine the target operating current and target operating gain corresponding to each channel to be tested, so that each channel to be tested performs fluorescence detection with the target operating current and target operating gain.

[0050] It is understood that the target operating current refers to the operating current of the PCR device to be calibrated after calibration, and the target operating gain refers to the operating gain of the PCR device to be calibrated after calibration. The PCR device to be calibrated can detect the fluorescent product according to the target operating current and target operating gain. The processor can adjust the operating current of the PCR device to be calibrated according to the current adjustment mode, or adjust the operating gain according to the gain adjustment mode, or simultaneously enter the current and gain adjustment modes to adjust the current and gain, so as to determine the operating current and operating gain after adjustment, which are used as the target operating current and target operating gain of the PCR device to be calibrated, so that each channel to be measured performs fluorescence detection at the target operating current and target operating gain.

[0051] In an embodiment of the present application, determining the target operating current and / or target operating gain of the PCR device to be calibrated includes: determining the deviation percentage between the first fluorescence data detected in real time and the standard fluorescence data; when the deviation percentage is within a preset deviation percentage range, determining the current operating current and operating gain as the target operating current and target operating gain, respectively; when the deviation percentage is not within the preset deviation percentage range, continuing to adjust the operating current and / or operating gain until the deviation percentage is within the preset deviation percentage range.

[0052] It will be understood that the deviation percentage refers to the ratio of the difference between the first fluorescence data and the standard fluorescence data to the standard fluorescence data. The preset deviation percentage range is an error range pre-set by the technician based on the deviation percentage. For each channel to be measured, while adjusting the operating current and / or operating gain according to the adjustment mode, the processor needs to determine in real time the first fluorescence data of the fluorescent substance at the measured well position based on the adjusted operating current and / or operating gain, and then detect the difference between the real-time detected first fluorescence data and the standard fluorescence data to obtain the deviation percentage between the two. If the deviation percentage is within the preset deviation percentage range, it indicates that the operating current and / or operating gain have approached the usage standard, and adjustment can be stopped, and the current operating current and operating gain can be determined as the target operating current and target operating gain, respectively. If the deviation percentage is not within the preset deviation percentage range, the operating current and / or operating gain can be adjusted again until the deviation percentage is within the preset deviation percentage range. For example, the preset deviation percentage range can be selected from ±1% to ±5%, and preferably, the preset deviation percentage range can be ±2%.

[0053] In an embodiment of the present application, the method also includes: obtaining a first adjustable range of the operating gain and / or a second adjustable range of the operating current; for each channel to be tested, when the operating gain exceeds the first adjustable range and / or the operating current exceeds the second adjustable range, outputting a calibration failure message.

[0054] It is understood that the first adjustable range of the operating gain refers to the interval between the upper and lower limits of the operating gain adjustment, and the second adjustable range of the operating current refers to the interval between the upper and lower limits of the operating current adjustment. The first adjustable range and the second adjustable range are adjustment ranges limited by hardware. If the first or second adjustable range is exceeded, and the deviation percentage between the first fluorescence data and the standard fluorescence data measured by the calibration fluorescent product is still not within the preset deviation percentage range, it indicates that the PCR device itself has a fault and requires repair, so the processor outputs a calibration failure message.

[0055] In an embodiment of the present application, for each channel to be tested, the operating current and / or operating gain of the PCR device to be calibrated are adjusted according to the adjustment mode to determine the target operating current and target operating gain of the PCR device to be calibrated, including: for each channel to be tested, after each adjustment of the operating current and / or operating gain according to the adjustment mode and the adjustment ratio, the deviation percentage between the first fluorescence data detected after each adjustment and the standard fluorescence data is determined; for each channel to be tested, if the corresponding deviation percentage after the last adjustment is within a preset deviation percentage range, the current operating current and operating gain are determined as the target operating current and target operating gain of the channel to be tested; for each channel to be tested, if the corresponding deviation percentage after the last adjustment is not within the preset deviation percentage range, the current operating current and / or operating gain continue to be adjusted according to the first fluorescence data detected after the last adjustment and the adjustment ratio until the deviation percentage between the current first fluorescence data and the standard fluorescence data is within the preset deviation percentage range or the number of adjustments is greater than or equal to the preset number threshold.

[0056] Referring to Figure 3, for the calibration process of the standard (calibration fluorescent substance), after the instrument to be calibrated (PCR device to be calibrated) starts the instrument calibration mode, the PC opens the calibration software and connects to the instrument to be calibrated, and the standard value of the standard (standard fluorescence data) is input from the PC. Select N channel, insert the standard into the selected test well position, initialize the current and gain value of N channel, and control the instrument to perform fluorescence acquisition. For each channel to be measured, the processor can adjust the operating current and / or operating gain multiple times according to the adjustment ratio. Before each adjustment, it is necessary to determine whether the current number of adjustments is greater than or equal to the maximum number of adjustments (adjustment number threshold). The preset number threshold is the adjustment number threshold set in advance by the technician to limit the calibration time. If the current number of adjustments is less than the maximum number of adjustments, then after each adjustment of the operating current, the first fluorescence data of the calibration fluorescent substance is re-detected according to the adjusted operating current, so that the ratio between the first fluorescence data and the standard fluorescence data can be re-determined. Specifically, the adjustment can be performed according to a linear adjustment ratio. At the same time, the deviation percentage between the first fluorescence data and the standard fluorescence data can be determined after each adjustment. In the gain adjustment mode, the operating gain is adjusted according to the adjustment ratio, and the ratio and deviation percentage between the first fluorescence data and the standard fluorescence data can be re-determined according to the above method. Furthermore, if the corresponding ratio after each adjustment is within the preset ratio range, the processor can determine that the corresponding adjustment mode after the current adjustment is the current adjustment mode, i.e., the next adjustment mode is the current adjustment mode. If the corresponding ratio after each adjustment is within the preset ratio range, the next adjustment mode is determined to be the gain adjustment mode, or the gain adjustment mode and the current adjustment mode. If the corresponding deviation percentage after the previous adjustment is within the preset deviation percentage range (set threshold), the current operating current and operating gain can be determined as the target operating current and target operating gain of the channel to be measured. If the corresponding deviation percentage after the previous adjustment is not within the preset deviation percentage range, the current operating current and / or operating gain are continuously adjusted according to the first fluorescence data detected after the previous adjustment and the adjustment ratio until the deviation percentage between the current first fluorescence data and the standard fluorescence data is within the preset deviation percentage range, or the number of adjustments exceeds the maximum number of adjustments. If the deviation percentage between the first fluorescence data and the standard fluorescence data is still not within the preset deviation percentage range after the maximum number of adjustments, a calibration failure message is output. If the deviation percentage between the first fluorescence data and the standard fluorescence data falls within the preset deviation percentage range within the maximum number of adjustments, a calibration success message is output. If calibration of all channels is complete, the calibration process ends. If not, calibration proceeds to the next channel, and the above process is repeated until all channels are calibrated.

[0057] In an embodiment of the present application, if during the adjustment process, its operating current or operating gain reaches a preset upper limit or lower limit, and the deviation percentage between the first fluorescence data and the standard fluorescence data is not within the preset deviation percentage range, the processor can control the PCR device to be calibrated to stop calibration and output a calibration failure message.

[0058] In an embodiment of the present application, the method further includes: for each channel to be tested, if the deviation percentage between the current first fluorescence data and the standard fluorescence data is not within a preset deviation percentage range and the number of adjustments is not within a preset number threshold, outputting a calibration failure message.

[0059] Specifically, if the number of adjustments exceeds a preset threshold, and the deviation percentage between the current first fluorescence data and the standard fluorescence data is not within a preset deviation percentage range, a calibration failure message may be output, and the calibration operation may be terminated. Alternatively, if during the adjustment process, the operating current and operating gain reach the upper or lower limit of the hardware limitations, and the deviation percentage between the first fluorescence data and the standard fluorescence data is not within the preset deviation percentage range, the processor may control the PCR device to be calibrated to terminate calibration and output a calibration failure message.

[0060] Through the above scheme, the ratio between the actual value and the measured value of the calibration fluorescence is determined by testing the well positions to determine the current adjustment mode or gain adjustment mode, thereby adjusting the operating current and / or operating gain of the PCR device to be calibrated. Each channel to be tested performs fluorescence detection based on the adjusted target operating current and target operating gain. The operating parameters of a multi-channel PCR device can be adjusted in different modes, ensuring that the adjusted PCR device meets the operating standards and the fluorescence performance requirements of the instrument manufacturer, thereby improving the accuracy of PCR device detection.

[0061] FIG1 is a schematic flow diagram of a calibration method for a PCR device in one embodiment. It should be understood that, although the various steps in the flow diagram of FIG1 are shown sequentially as indicated by the arrows, these steps are not necessarily performed sequentially in the order indicated by the arrows. Unless otherwise specified herein, there is no strict order restriction on the execution of these steps, and these steps may be performed in other orders. Furthermore, at least a portion of the steps in FIG1 may include multiple sub-steps or multiple stages, and these sub-steps or stages are not necessarily performed at the same time, but may be performed at different times. The execution order of these sub-steps or stages is not necessarily sequential, but may be performed in rotation or alternating with other steps or at least a portion of the sub-steps or stages of other steps.

[0062] FIG4 schematically shows a block diagram of a PCR device according to an embodiment of the present application. The present application also provides a PCR device 400, which may include:

[0063] Hole position to be measured 410;

[0064] At least one channel to be tested 420;

[0065] And a calibration device 500 for PCR equipment, used to perform the above calibration method for PCR equipment.

[0066] FIG5 schematically shows a block diagram of a calibration device for a PCR device according to an embodiment of the present application. As shown in FIG5 , the present application embodiment provides a calibration device 500 for a PCR device, which may include:

[0067] Memory 510 configured to store instructions; and

[0068] The processor 520 is configured to call instructions from the memory 510 and implement the above-mentioned calibration method for the PCR device when executing the instructions.

[0069] Specifically, in the embodiment of the present application, the processor 520 may be configured to:

[0070] Controlling each channel to be measured to detect the calibration fluorescent material of the well position to be measured to obtain first fluorescence data of the well position to be measured;

[0071] Obtain standard fluorescence data of calibration fluorophores;

[0072] determining a ratio between the first fluorescence data and the standard fluorescence data;

[0073] determining an adjustment mode of the PCR device to be calibrated according to the ratio, the adjustment mode being a current adjustment mode and / or a gain adjustment mode;

[0074] The operating current and / or operating gain of the PCR device to be calibrated is adjusted according to the adjustment mode to determine the target operating current and target operating gain corresponding to each channel to be tested, so that each channel to be tested performs fluorescence detection with the target operating current and target operating gain.

[0075] In the embodiment of the present application, the processor 520 may also be configured to:

[0076] Determining the adjustment mode of the PCR device to be calibrated according to the ratio includes: when the ratio is within the preset ratio range, determining the adjustment mode to be the current adjustment mode; when the ratio is not within the preset ratio range and the working gain is continuously adjusted, determining the adjustment mode to be the gain adjustment mode.

[0077] In the embodiment of the present application, the processor 520 may also be configured to:

[0078] Determining the adjustment mode of the PCR device to be calibrated according to the ratio also includes: when the ratio is not within a preset ratio range and the working gain is not continuously adjusted, determining the adjustment mode to be the current adjustment mode and the gain adjustment mode.

[0079] In an embodiment of the present application, determining the target operating current and / or target operating gain of the PCR device to be calibrated includes: determining the deviation percentage between the first fluorescence data detected in real time and the standard fluorescence data; when the deviation percentage is within a preset deviation percentage range, determining the current operating current and operating gain as the target operating current and target operating gain, respectively; when the deviation percentage is not within the preset deviation percentage range, continuing to adjust the operating current and / or operating gain until the deviation percentage is within the preset deviation percentage range.

[0080] In the embodiment of the present application, the processor 520 may also be configured to:

[0081] The method further includes: obtaining a first adjustable range of the operating gain and a second adjustable range of the operating current; and outputting calibration failure information when the operating gain exceeds the first adjustable range and / or the operating current exceeds the second adjustable range.

[0082] In the embodiment of the present application, the processor 520 may also be configured to:

[0083] The method further includes: after determining the target operating current and target operating gain corresponding to the current channel to be tested, determining the target operating current and target operating gain corresponding to the next channel to be tested, until all channels to be tested are calibrated.

[0084] Through the above scheme, by detecting the hole position to be measured, the ratio between the actual value and the measured value of the calibration fluorescent product is obtained to determine the current regulation mode or the gain regulation mode, or by jointly regulating the current and gain regulation modes, the operating current and the operating gain of the PCR device to be calibrated are adjusted. In addition, after the target operating current and the target operating gain are determined for all the channels to be measured, it is determined that the calibration of the PCR device to be calibrated is complete. Then, each channel to be measured can perform fluorescence detection based on the adjusted target operating current and target operating gain. Therefore, the embodiment of the present application can be used for multi-channel PCR equipment to perform linear proportional adjustment of operating parameters in different modes. The PCR device after adjustment is made to meet the use standards, meet the fluorescence performance requirements of the instrument production factory, and improve the accuracy of PCR device detection.

[0085] Figure 6 schematically shows a schematic diagram of a calibration system for PCR equipment according to an embodiment of the present application. As shown in Figure 6, the embodiment of the present application provides a calibration system for PCR equipment, which may include a PCR device to be calibrated 610 and a calibration device 500 for the PCR device. Specifically, the calibration device is a PC terminal, and the PC terminal and the PCR device to be calibrated are controlled by communication. The above-mentioned method for PCR equipment is implemented by the calibration software on the PC terminal. Among them, the calibration software on the PC terminal includes a first communication module, an inlet and outlet control module, a first calibration module and a second calibration module. Specifically, the first communication module can realize communication drive, complete the communication function with the PCR device to be calibrated, and control the receiving instrument data. The inlet and outlet control module selects a channel for calibration analysis, automatically inserts the calibration fluorescent product into the well position to be measured through the software, and sets the standard fluorescence data value on the PC terminal software. The first calibration module collects fluorescence data for the well position to be measured, adjusts the LED current and the op amp gain, so that the collected fluorescence data reaches within the preset range. The second calibration module uses linear proportional calibration. After completing the current and gain calibration at the well position to be measured, the calibrated target working current and target working gain are used to collect fluorescence data of all fluxes and report them to the PC. The PCR device to be calibrated includes a second communication module and a calibration analysis module. The second communication module can enter the instrument calibration mode through the interface button function and communicate with the PC software through the network or other communication drivers. The calibration analysis module can send a command through communication on the PC to start the calibration of the device. After receiving the command, the instrument to be calibrated starts working and reports the fluorescence collection data to the PC, waiting for the PC to send the target working current and target working gain. After receiving the parameters, the instrument will immediately take effect on the parameters and upload the calibrated fluorescence data to the PC software in real time.

[0086] An embodiment of the present application further provides a machine-readable storage medium having stored thereon instructions for causing a machine to execute the above-mentioned calibration method for a PCR device.

[0087] In one embodiment, a computer device is provided, which can be a server, and its internal structure diagram can be shown in Figure 7. The computer device includes a processor A01, a network interface A02, a memory (not shown) and a database (not shown) connected by a system bus. Among them, the processor A01 of the computer device is used to provide computing and control capabilities. The memory of the computer device includes an internal memory A03 and a non-volatile storage medium A04. The non-volatile storage medium A04 stores an operating system B01, a computer program B02 and a database (not shown). The internal memory A03 provides an environment for the operation of the operating system B01 and the computer program B02 in the non-volatile storage medium A04. The database of the computer device is used to store calibration data for PCR equipment. The network interface A02 of the computer device is used to communicate with an external terminal via a network connection. When the computer program B02 is executed by the processor A01, a calibration method for PCR equipment is implemented.

[0088] Those skilled in the art will understand that the structure shown in FIG7 is merely a block diagram of a portion of the structure related to the solution of the present application, and does not constitute a limitation on the computer device to which the solution of the present application is applied. The specific computer device may include more or fewer components than shown in the figure, or combine certain components, or have a different arrangement of components.

[0089] Those skilled in the art will appreciate that the embodiments of the present application can be provided as methods, systems or computer program products. Therefore, the present application can adopt the form of a complete hardware embodiment, a complete software embodiment, or an embodiment in combination with software and hardware. Moreover, the present application can adopt the form of a computer program product implemented on one or more computer-usable storage media (including but not limited to magnetic disk storage, CD-ROM, optical storage, etc.) that contain computer-usable program code.

[0090] The present application is described with reference to the flowcharts and / or block diagrams of the methods, devices (systems), and computer program products according to the embodiments of the present application. It should be understood that each process and / or box in the flowchart and / or block diagram, as well as the combination of the processes and / or boxes in the flowchart and / or block diagram, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general-purpose computer, a special-purpose computer, an embedded processor, or other programmable data processing device to produce a machine, so that the instructions executed by the processor of the computer or other programmable data processing device produce a device for implementing the functions specified in one or more processes in the flowchart and / or one or more boxes in the block diagram.

[0091] These computer program instructions may also be stored in a computer-readable memory that can direct a computer or other programmable data processing device to operate in a specific manner, so that the instructions stored in the computer-readable memory produce a product including an instruction device that implements the functions specified in one or more processes in the flowchart and / or one or more boxes in the block diagram.

[0092] These computer program instructions can also be loaded onto a computer or other programmable data processing device so that a series of operating steps are executed on the computer or other programmable device to produce a computer-implemented process, so that the instructions executed on the computer or other programmable device provide steps for implementing the functions specified in one or more processes in the flowchart and / or one or more boxes in the block diagram.

[0093] In a typical configuration, a computing device includes one or more processors (CPUs), input / output interfaces, network interfaces, and memory.

[0094] The memory may include non-permanent memory in a computer-readable medium, random access memory (RAM) and / or non-volatile memory in the form of read-only memory (ROM) or flash RAM. The memory is an example of a computer-readable medium.

[0095] Computer-readable media includes permanent and non-permanent, removable and non-removable media that can be implemented by any method or technology to store information. The information can be computer-readable instructions, data structures, program modules or other data. Examples of computer storage media include, but are not limited to, phase change memory (PRAM), static random access memory (SRAM), dynamic random access memory (DRAM), other types of random access memory (RAM), read-only memory (ROM), electrically erasable programmable read-only memory (EEPROM), flash memory or other memory technology, compact disc read-only memory (CD-ROM), digital versatile disc (DVD) or other optical storage, magnetic cassettes, magnetic disk storage or other magnetic storage devices or any other non-transmission media that can be used to store information that can be accessed by a computing device. As defined herein, computer-readable media does not include transitory computer-readable media (transitory media), such as modulated data signals and carrier waves.

[0096] It should also be noted that the terms "comprises," "includes," or any other variations thereof are intended to encompass non-exclusive inclusion, such that a process, method, commodity, or apparatus that includes a series of elements includes not only those elements but also other elements not explicitly listed, or includes elements inherent to such process, method, commodity, or apparatus. In the absence of further limitations, an element defined by the phrase "comprises a ..." does not exclude the presence of other identical elements in the process, method, commodity, or apparatus that includes the element.

[0097] The above are merely embodiments of the present application and are not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present application should all be included within the scope of the claims of the present application.

Claims

1. A calibration method for a PCR device, characterized in that, The PCR device to be calibrated includes a to-be-tested well position and at least one to-be-tested channel, and the calibration method includes: Controlling each to-be-tested channel to detect the calibration fluorophore of the to-be-tested well position to obtain the first fluorescence data of the to-be-tested well position; Obtaining the standard fluorescence data of the calibration fluorophore; Determining the ratio between the first fluorescence data and the standard fluorescence data; Determining the adjustment mode of the PCR device to be calibrated according to the ratio, where the adjustment mode is a current adjustment mode and / or a gain adjustment mode; Adjusting the working current and / or the working gain of the PCR device to be calibrated according to the adjustment mode to determine the target working current and the target working gain corresponding to each to-be-tested channel, so that each to-be-tested channel performs fluorescence detection with the target working current and the target working gain.

2. The calibration method for a PCR device according to claim 1, characterized in that, The determining the adjustment mode of the PCR device to be calibrated according to the ratio includes: When the ratio is within a preset ratio range, determining the adjustment mode as the current adjustment mode; When the ratio is not within the preset ratio range and the working gain is continuously adjustable, determining the adjustment mode as the gain adjustment mode.

3. The calibration method for a PCR device according to claim 2, wherein, The determining the adjustment mode of the PCR device to be calibrated according to the ratio further includes: When the ratio is not within the preset ratio range and the working gain is not continuously adjustable, determining the adjustment mode as the current adjustment mode and the gain adjustment mode.

4. The calibration method for a PCR device according to claim 1, characterized in that, Determining the target working current and the target working gain of the PCR device to be calibrated includes: Determining the deviation percentage between the first fluorescence data detected in real time and the standard fluorescence data; When the deviation percentage is within a preset deviation percentage range, respectively determining the current working current and the working gain as the target working current and the target working gain; When the deviation percentage is not within the preset deviation percentage range, continuing to adjust the working current and / or the working gain until the deviation percentage is within the preset deviation percentage range.

5. The calibration method for a PCR device according to claim 1, characterized in that, The method further includes: Obtaining the first adjustable range of the working gain and / or the second adjustable range of the working current; When the working gain exceeds the first adjustable range and / or the working current exceeds the second adjustable range, outputting a calibration failure message.

6. The calibration method for a PCR device according to claim 1, wherein, The method further includes: After determining the target working current and the target working gain corresponding to the current to-be-tested channel, determining the target working current and the target working gain corresponding to the next to-be-tested channel until the calibration of all to-be-tested channels is completed.

7. A calibration device for a PCR device, characterized in that, Includes: A memory configured to store instructions; A processor configured to call the instructions from the memory and, when executing the instructions, be capable of implementing the calibration method for a PCR device according to any one of claims 1 to 6.

8. A PCR device, characterized in that, Includes: A to-be-tested well position; At least one to-be-tested channel; A calibration device for a PCR device according to claim 7.

9. A calibration system for a PCR device, characterized in that, Includes: A PCR device to be calibrated, including a to-be-tested well position and at least one to-be-tested channel; A calibration device for a PCR device according to claim 8.

10. A machine-readable storage medium, characterized in that, Instructions are stored on the machine-readable storage medium, and the instructions are used to cause the machine to execute the calibration method for a PCR device according to any one of claims 1 to 6.

Images