Calibration method, device and system for PCR (Polymerase Chain Reaction) equipment and PCR equipment

Abstract

The invention discloses a calibration method, device and system for PCR equipment and the PCR equipment. The calibration method comprises the following steps: controlling a to-be-detected channel to detect a calibration fluorescent product of a to-be-detected hole site so as to obtain first fluorescence data of the to-be-detected hole site; acquiring standard fluorescence data of the calibration fluorescent product; determining a ratio between the first fluorescence data and the standard fluorescence data; determining an adjustment mode of the to-be-calibrated PCR device according to the proportion, the adjustment mode being a current adjustment mode and / or a gain adjustment mode; and adjusting the working current and / or the working gain of the to-be-calibrated PCR equipment according to the adjustment mode to determine the target working current and the target working gain of the to-be-calibrated PCR equipment, so that each to-be-detected channel performs fluorescence detection with the target working current and the target working gain. According to the invention, the working parameters of the multichannel PCR equipment can be adjusted in different modes, so that the detection accuracy of the PCR equipment is improved.

Description

Technical Field

[0001] The present application relates to the technical field of fluorescence detection, and specifically relates to a calibration method, device, system, PCR device and storage medium for a PCR device. Background Art

[0002] A real-time fluorescence quantitative PCR instrument (POCT) is used to detect the fluorescence in the cycling process, and the fluorescence data is collected by a computer connected to the real-time device. Among them, the data is displayed in the form of a standard curve through 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 the specific DNA sequence in the sample to be tested. 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 the calibration of the instrument parameters before the gene amplification reaction and the 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 acquisition process. If the instrument performance is unqualified, it will lead to inaccurate measurement and have an adverse impact on scientific research. Summary of the Invention

[0003] The purpose of the embodiments of the present application is to provide a calibration method, device, system, PCR device and storage medium for a PCR device, so as to solve the technical problem of inaccurate measurement of PCR instruments in the prior art.

[0004] The first aspect of the present application provides a calibration method for a PCR device. The PCR device to be calibrated includes a to-be-tested well position and at least one to-be-tested channel. The calibration method includes:

[0005] Controlling each to-be-tested channel to detect the calibration fluorophore in the to-be-tested well position to obtain the first fluorescence data of the to-be-tested well position;

[0006] Obtaining the standard fluorescence data of the calibration fluorophore;

[0007] Determining the ratio between the first fluorescence data and the standard fluorescence data;

[0008] Determining the adjustment mode of the PCR device to be calibrated according to the ratio. The adjustment mode is a current adjustment mode and / or a gain adjustment mode;

[0009] Adjusting the working current and / or working gain of the PCR device to be calibrated according to the adjustment mode to determine the target working current and 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 target working gain.

[0010] In an embodiment of the present application, 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.

[0011] In an embodiment of the present application, 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.

[0012] In an embodiment of the present application, determining the target working current and 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, determining the current working current and working gain as the target working current and target working gain respectively; when the deviation percentage is not within the preset deviation percentage range, continue to adjust the working current and / or working gain until the deviation percentage is within the preset deviation percentage range.

[0013] In an embodiment of the present application, 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.

[0014] In an embodiment of the present application, the method further includes: after determining the target working current and target working gain corresponding to the current channel to be tested, determining the target working current and target working gain corresponding to the next channel to be tested until the calibration of all channels to be tested is completed.

[0015] A second aspect of the present application provides a calibration device for a PCR device, including:

[0016] A memory configured to store instructions; and

[0017] A processor configured to call instructions from the memory and capable of implementing the calibration method for the PCR device according to the above.

[0018] A third aspect of the present application provides a PCR device, including:

[0019] Wells to be tested;

[0020] At least one channel to be tested; and

[0021] The calibration device for the PCR device according to the above.

[0022] A fourth aspect of the present application provides a calibration system for a PCR device, including

[0023] a PCR device to be calibrated, including a to-be-tested well position and at least one to-be-tested channel; and

[0024] the above-mentioned calibration device for a PCR device

[0025] A fifth aspect of the present application provides a machine-readable storage medium, characterized in that instructions are stored on the machine-readable storage medium, and the instructions are used to cause a machine to execute the calibration method for a PCR device according to the above.

[0026] Through the above solution, by detecting the target well positions selected from the to-be-tested well positions, the ratio between the actual value and the measured value of the calibration fluorophore is obtained to determine the current adjustment mode or the gain adjustment mode, so as to adjust the working current and / or the working gain of the PCR device to be calibrated. Each to-be-tested channel performs fluorescence detection based on the adjusted target working current and target working gain. The present application can adjust the working parameters of single-flux and multi-flux multi-channel PCR devices in different modes, so that the adjusted PCR device meets the usage standards, meets the fluorescence performance requirements for instrument production and factory shipment, and improves the accuracy of PCR device detection.

[0027] Other features and advantages of the embodiments of the present application will be described in detail in the subsequent specific implementation section. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The drawings are used to provide a further understanding of the embodiments of the present application, and constitute a part of the specification. Together with the following specific implementation, they are used to explain the embodiments of the present application, but do not constitute a limitation to the embodiments of the present application. In the drawings:

[0029] Figure 1 Schematically shows a flowchart of a calibration method for a PCR device according to an embodiment of the present application;

[0030] Figure 2 Schematically shows a circuit diagram of current adjustment and gain adjustment according to an embodiment of the present application;

[0031] Figure 3 Schematically shows a flowchart of a calibration method for a PCR device according to a specific embodiment of the present application;

[0032] Figure 4 Schematically shows a structural block diagram of a PCR device according to an embodiment of the present application;

[0033] Figure 5 Schematically shows a structural block diagram of a calibration device for a PCR device according to an embodiment of the present application;

[0034] Figure 6 Schematically shown is a schematic diagram of a calibration system for a PCR device according to an embodiment of the present application;

[0035] Figure 7 Schematically shown is a schematic structural diagram of a computer device according to an embodiment of the present application. Detailed implementation manners

[0036] To make the objectives, 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 with reference to the accompanying drawings in the embodiments of the present application. It should be understood that the specific implementation manners described herein are only for explaining and illustrating the embodiments of the present application, and are not used to limit the embodiments of the present application. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present application without creative efforts belong to the scope protected by the present application.

[0037] It should be noted that if there are directional indications (such as up, down, left, right, front, back,...) involved in the embodiments of the present application, the directional indications are only used to explain the relative positional relationship and movement conditions between components in a specific posture (as shown in the drawings). If the specific posture changes, the directional indications will also change accordingly.

[0038] In addition, if there are descriptions such as "first" and "second" involved 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 quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include at least one such feature. In addition, the technical solutions between various embodiments may be combined with each other, but it must be based on the fact that those of ordinary skill in the art can implement them. When the combination of technical solutions results in contradictions or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection required by the present application.

[0039] Figure 1 Schematically shown is a flowchart of a calibration method for a PCR device according to an embodiment of the present application. As Figure 1 shown, the embodiments of the present application provide a calibration method for a PCR device, and the calibration method may include the following steps.

[0040] The PCR device to be calibrated includes a to-be-tested well position and at least one to-be-tested channel. It can be understood that the PCR device, also known as a PCR amplifier, is an instrument that amplifies specific DNA using the PCR technique. The PCR device to be calibrated refers to a PCR device whose fluorescence acquisition value has not been calibrated to meet the usage standard. The PCR device to be calibrated can include at least one to-be-tested well position and at least one to-be-tested channel. Understandably, a well position refers to the detection unit of the PCR device for a fluorescence sample, usually referring to the well position of the PCR device for placing the fluorescence sample and the reaction solution. Each well position can perform an independent PCR reaction, and simultaneous detection of multiple samples can be achieved through the design of multiple well positions. Among them, the throughput of different well positions can be different. Throughput refers to the number of samples that can be detected in one PCR reaction. The higher the throughput, the more samples can be detected in one PCR reaction. A channel refers to the detection channel of the PCR device for a fluorescence sample, usually referring to the channel of the PCR device for detecting fluorescence signals. The fluorescence products of different well positions can be detected under the same channel. The to-be-tested well position refers to the well position to be calibrated, and the to-be-tested channel refers to the channel to be calibrated.

[0041] Then, during the calibration process of the PCR device, first, any one of the to-be-tested well positions can be selected as the target well position from multiple to-be-tested well positions. Specifically, if the PCR device to be calibrated is a single-well PCR instrument, the only to-be-tested well position of the PCR device to be calibrated is the target well position. If the PCR device to be calibrated is a multi-well PCR instrument, any one of the to-be-tested well positions can be selected as the target well position.

[0042] S102, Control each to-be-tested channel to detect the calibration fluorescence product of the to-be-tested well position to obtain the first fluorescence data of the to-be-tested well position.

[0043] S104, Obtain the standard fluorescence data of the calibration fluorescence product.

[0044] It can be understood that the calibration fluorescence product, that is, the fluorescence calibration product, usually refers to a standard sample with known fluorescence characteristics, which can be used as a calibration fluorescence product for calibrating fluorescence detection instruments or methods. The first fluorescence data refers to the fluorescence characteristic data of the calibration fluorescence product tested at different wavelengths, including but not limited to fluorescence intensity, emission rate, etc. The standard fluorescence data refers to the known standard fluorescence data of the calibration fluorescence product. Specifically, the processor can control the to-be-tested channel to detect the calibration fluorescence product of the to-be-tested well position to obtain the first fluorescence data of the calibration fluorescence product of the to-be-tested target well position under the detection of the to-be-tested channel.

[0045] S106, Determine the ratio between the first fluorescence data and the standard fluorescence data.

[0046] Specifically, during the fluorescence calibration process, the ratio between the first fluorescence data and the standard fluorescence data can be determined, so as to compare the first fluorescence data and the standard fluorescence data of the measured calibration fluorophore, and determine whether the test performance of the instrument is accurate.

[0047] S108. Determine the adjustment mode of the PCR device to be calibrated according to the ratio. The adjustment mode is a current adjustment mode and / or a gain adjustment mode.

[0048] It can be understood that the current adjustment mode means that by adjusting the current of the PCR device to be calibrated, the reaction temperature and reaction speed of the PCR reaction can be controlled. During the PCR reaction process, different temperatures will affect processes such as DNA denaturation, annealing, and extension. Adjusting the current can affect the amplification efficiency and quality of DNA, thereby affecting the measured fluorescence data results. The gain adjustment mode means that by adjusting the operational amplifier gain of the PCR device to be calibrated, the sensitivity and noise level of the detection system can be controlled, thereby affecting the measured fluorescence data results. For different channels, different fluorescence labels can be detected. The ratio between the true data and the test data of the fluorescence 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 is adjusted through the current adjustment mode or the gain adjustment mode, or adjusted according to both the current adjustment mode and the gain adjustment mode according to the ratio between the first fluorescence data and the standard fluorescence data. Specifically, referring to Figure 2 , the current adjustment mode means adjusting the current of the LED lamp at the emission end of the PCR device to adjust the light emission intensity of the LED lamp. The gain adjustment mode means adjusting the signal at the receiving end of the PCR device and performing gain adjustment on the signal.

[0049] In the embodiments of the present application, 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 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.

[0050] In the embodiments of the present application, 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.

[0051] It can be understood that the working current refers to the current at which the PCR device to be calibrated detects the calibration fluorophore. The working gain refers to the operational amplifier gain at which the PCR device to be calibrated detects the calibration fluorophore. When the fluorophores at different wells to be measured are detected simultaneously through the channel to be measured, the working current and the working gain of the wells to be measured are the same. The preset ratio range refers to the ratio range preset by technicians according to technical experience. For example, when the ratio is within [1 / 2, 2], the current adjustment mode is adopted to adjust the current of the LED at the transmitting end. When the ratio is not within [1 / 2, 2], the gain adjustment mode is adopted to adjust the gain of the received signal, or the current and the gain are adjusted simultaneously.

[0052] Among them, if the ratio is not within the preset ratio range, in the case where the working gain can be continuously adjusted, only the gain can be adjusted for rough adjustment until the ratio is within the preset ratio range. If the working gain cannot be continuously adjusted, for example, each adjustment is increased or decreased at intervals, then there may be a situation where the ratio of the previous adjustment is less than 1 / 2 and the ratio of the next adjustment is greater than 2, and accurate adjustment can never be achieved. Then, in this case, the current adjustment mode can be entered, and the current is adjusted until the ratio is within the preset ratio range. When the ratio is not within the preset ratio range, it can be adjusted either through the gain adjustment mode or through the current adjustment mode.

[0053] In the embodiments of the present application, for each channel to be measured, determining the adjustment mode of the PCR device to be calibrated according to the ratio includes: for each channel to be measured, obtaining the adjustment ratio for adjusting the working current and / or the working gain multiple times; for each channel to be measured, after adjusting the working current and / or the working gain each time according to the adjustment mode and the adjustment ratio, re-determining the ratio between the first fluorescence data and the standard fluorescence data detected after each adjustment; for each channel to be measured, when the ratio corresponding to each adjustment is within the preset ratio range, determining the adjustment mode corresponding to each adjustment as the current adjustment mode; for each channel to be measured, when the ratio corresponding to each adjustment is within the preset ratio range, determining the adjustment mode corresponding to each adjustment as the gain adjustment mode, or as the gain adjustment mode and the current adjustment mode.

[0054] Specifically, for each channel to be measured, the processor can adjust the working current and working gain multiple times according to an adjustment ratio. The adjustment ratio can be a linear ratio or a non-linear ratio. In the current adjustment mode, the working current is adjusted according to the adjustment ratio. Specifically, it can be based on the previous working current, and the set adjustment ratio is added or subtracted to obtain the current working current. After each adjustment of the working current, the first fluorescence data of the fluorescent product is re-detected and calibrated according to the adjusted working current, so that the ratio between the first fluorescence data and the standard fluorescence data can be re-determined. In the gain adjustment mode, the working 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 method. Specifically, it can be based on the previous working gain, and the set adjustment ratio is added or subtracted to obtain the current working gain. Among them, the adjustment ratio of the working current can be the first adjustment ratio, and the adjustment ratio of the working gain can be the second adjustment ratio. Further, when the ratio corresponding to each adjustment is within the preset ratio range, the processor can determine that the adjustment mode corresponding to this adjustment is the current adjustment mode, that is, the next adjustment mode is the current adjustment mode. When the ratio corresponding to each adjustment is within the preset ratio range, it is determined that the next adjustment mode is the gain adjustment mode. The preset ratio range refers to the ratio range preset by technicians according to technical experience. For example, when the ratio corresponding to the adjustment is within [1 / 2, 2], a linear adjustment method is used to adjust the current. When the ratio corresponding to the adjustment is not within [1 / 2, 2], a linear adjustment method is used to adjust the gain.

[0055] S110. Adjust the working current and / or working gain of the PCR device to be calibrated according to the adjustment mode to determine the target working current and target working gain corresponding to each channel to be measured, so that each channel to be measured performs fluorescence detection with the target working current and target working gain.

[0056] It can be understood that the target working current refers to the working current after the PCR device to be calibrated is calibrated, and the target working gain refers to the working gain after the PCR device to be calibrated is calibrated. The PCR device to be calibrated can detect the fluorescent product according to the target working current and target working gain. The processor can adjust the working current of the PCR device to be calibrated according to the current adjustment mode, or adjust the working gain according to the gain adjustment mode, or enter the current and gain adjustment modes simultaneously to adjust the current and gain to determine the working current and working gain after adjustment as the target working current and target working gain of the PCR device to be calibrated, so that each channel to be measured performs fluorescence detection with the target working current and target working gain.

[0057] In an embodiment of the present application, determining the target working current and / or target working gain of the PCR device to be calibrated includes: determining the percentage deviation between the first fluorescence data detected in real time and the standard fluorescence data; when the percentage deviation is within the preset percentage deviation range, determining the current working current and working gain as the target working current and target working gain respectively; when the percentage deviation is not within the preset percentage deviation range, continue to adjust the working current and / or working gain until the percentage deviation is within the preset percentage deviation range.

[0058] It can be understood that the percentage deviation refers to the ratio of the difference between the first fluorescence data and the standard fluorescence data to the standard fluorescence data. The preset percentage deviation range is an error range preset by those skilled in the art based on the percentage deviation. For each channel to be measured, during the process of adjusting the working current and / or working gain according to the adjustment mode, the processor needs to determine in real time the working current and / or working gain based on the adjustment to detect the first fluorescence data of the fluorescent product in the well to be measured in real time, so as to determine the difference between the first fluorescence data detected in real time and the standard fluorescence data, and obtain the percentage deviation between the two. If the percentage deviation is within the preset percentage deviation range, it means that the working current and / or working gain has approached the standard for use, and the adjustment can be stopped, and the current working current and working gain are determined as the target working current and target working gain respectively. If the percentage deviation is not within the preset percentage deviation range, the working current and / or working gain can be continued to be adjusted until the percentage deviation is within the preset percentage deviation range. For example, the preset percentage deviation range can be selected from ±1% to ±5%, preferably, the preset percentage deviation range can be ±2%.

[0059] In an embodiment of the present application, the method further includes: obtaining the first adjustable range of the working gain and / or the second adjustable range of the working current; for each channel to be measured, when the working gain exceeds the first adjustable range and / or the working current exceeds the second adjustable range, outputting a calibration failure message.

[0060] It can be understood that the first adjustable range of the working gain refers to the interval between the upper limit value and the lower limit value of the working gain adjustment, and the second adjustable range of the working current refers to the interval between the upper limit value and the lower limit value of the working current adjustment. Among them, the first adjustable range and the second adjustable range are the adjustment ranges limited by the hardware. If it exceeds the first adjustable range or the second adjustable range, and the percentage deviation between the first fluorescence data measured by the calibration fluorescent product and the standard fluorescence data is still not within the preset percentage deviation range, it means that the PCR device itself has a fault that needs to be repaired, so the processor outputs a calibration failure message.

[0061] In the embodiments of the present application, for each channel to be measured, adjusting the working current and / or working gain of the PCR device to be calibrated according to the adjustment mode to determine the target working current and target working gain of the PCR device to be calibrated includes: for each channel to be measured, after adjusting the working current and / or working gain according to the adjustment mode and adjustment ratio each time, determining the percentage deviation between the first fluorescence data detected after each adjustment and the standard fluorescence data; for each channel to be measured, when the percentage deviation corresponding to the previous adjustment is within the preset percentage deviation range, determining the current working current and working gain as the target working current and target working gain of the channel to be measured; for each channel to be measured, when the percentage deviation corresponding to the previous adjustment is not within the preset percentage deviation range, continuing to adjust the current working current and / or working gain according to the first fluorescence data detected after the previous adjustment and the adjustment ratio until the percentage deviation between the current first fluorescence data and the standard fluorescence data is within the preset percentage deviation range or the number of adjustment times is greater than or equal to the preset number threshold.

[0062] Reference Figure 3, for the calibration process of the standard product (calibration fluorescent product), after the instrument to be calibrated (PCR device to be calibrated) starts the instrument calibration mode, the calibration software is opened on the PC side and connected to the instrument to be calibrated, and the standard value (standard fluorescent data) of the standard product is input from the PC side. Select the N channel, insert the standard product into the selected test hole position, initialize the current and gain values of the N channel, and control the instrument to perform fluorescence acquisition. For each channel to be measured, the processor can adjust the working current and / or working 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 by technicians in advance 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 working current, the first fluorescence data of the calibration fluorescent product is re-detected according to the adjusted working current, so that the ratio between the first fluorescence data and the standard fluorescence data can be re-determined. Specifically, the adjustment can be made 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 working 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. Further, when the ratio corresponding to each adjustment is within the preset ratio range, the processor can determine that the adjustment mode corresponding to this adjustment is the current adjustment mode, that is, the next adjustment mode is the current adjustment mode. When the ratio corresponding to each adjustment is within the preset ratio range, it is determined that the next adjustment mode is the gain adjustment mode, or the gain adjustment mode and the current adjustment mode. If the deviation percentage corresponding to the previous adjustment is within the preset deviation percentage range (set threshold), the current working current and working gain can be determined as the target working current and target working gain of the channel to be measured. If the deviation percentage corresponding to the previous adjustment is not within the preset deviation percentage range, continue to adjust the current working current and / or working gain 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 is greater than the maximum number of adjustments. If the maximum number of adjustments is exceeded and the deviation percentage between the first fluorescence data and the standard fluorescence data is still not within the preset deviation percentage range, a calibration failure message is output. If the deviation percentage between the first fluorescence data and the standard fluorescence data is within the preset deviation percentage range during the adjustment within the maximum number of adjustments, a calibration success message is output. If the calibration of all channels to be measured is completed, the calibration process ends. If not, enter the calibration of the next channel to be measured and repeat the above process until the calibration of all channels to be measured is completed.

[0063] In another embodiment, if during the adjustment process, its working current or working gain reaches a preset upper or lower limit value, and the percentage deviation between the first fluorescence data and the standard fluorescence data is not within the preset percentage deviation range, the processor may control the PCR device to be calibrated to stop calibration and output a calibration failure message.

[0064] In an embodiment of the present application, the method further includes: for each channel to be measured, when the percentage deviation between the current first fluorescence data and the standard fluorescence data is not within the preset percentage deviation range, and the number of adjustment times is not within the preset number threshold, output a calibration failure message.

[0065] Specifically, if the number of adjustment times is greater than the preset number threshold, and the percentage deviation between the current first fluorescence data and the standard fluorescence data is not within the preset percentage deviation range, then a calibration failure message may be output and the calibration operation may be stopped. Or, if during the adjustment process, its working current and action gain reach the upper or lower limit value of the hardware limit, and the percentage deviation between the first fluorescence data and the standard fluorescence data is not within the preset percentage deviation range, the processor may control the PCR device to be calibrated to stop calibration and output a calibration failure message.

[0066] Through the above solution, by detecting the positions of the wells to be measured, the ratio between the actual value and the measured value of the calibration fluorophore is obtained to determine the current adjustment mode or the gain adjustment mode, thereby adjusting the working current and / or the working gain of the PCR device to be calibrated. Each channel to be measured performs fluorescence detection based on the adjusted target working current and target working gain. Different modes of adjustment can be performed on the working parameters of the multi-channel PCR device, so that the adjusted PCR device meets the usage standards and satisfies the fluorescence performance requirements for factory production of the instrument, improving the accuracy of PCR device detection.

[0067] Figure 4 A schematic block diagram of a PCR device according to an embodiment of the present application is shown. An embodiment of the present application further provides a PCR device 400, which may include:

[0068] Wells to be measured 410;

[0069] At least one channel to be measured 420;

[0070] And a calibration device 500 for the PCR device, which is used to execute the above-mentioned calibration method for the PCR device.

[0071] Figure 5 A schematic block diagram of a calibration device for a PCR device according to an embodiment of the present application is shown. As Figure 5 shown, an embodiment of the present application provides a calibration device 500 for a PCR device, which may include:

[0072] A memory 510, configured to store instructions; and

[0073] A processor 520, configured to call instructions from the memory 510 and capable of implementing the above method for controlling the boom when executing the instructions.

[0074] Specifically, in the embodiment of the present application, the processor 520 may be configured to:

[0075] Control each channel to be tested to detect the calibration fluorophore at the well position to be tested to obtain the first fluorescence data of the well position to be tested;

[0076] Obtain the standard fluorescence data of the calibration fluorophore;

[0077] Determine the ratio between the first fluorescence data and the standard fluorescence data;

[0078] Determine the adjustment mode of the PCR device to be calibrated according to the ratio, and the adjustment mode is a current adjustment mode and / or a gain adjustment mode;

[0079] Adjust 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 channel to be tested, so that each channel to be tested performs fluorescence detection with the target working current and the target working gain.

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

[0081] 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 that the adjustment mode is a current adjustment mode; when the ratio is not within the preset ratio range and the working gain is continuously adjustable, determining that the adjustment mode is a gain adjustment mode.

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

[0083] 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 that the adjustment mode is a current adjustment mode and a gain adjustment mode.

[0084] In an embodiment of the present application, determining the target working current and / or target working gain of the PCR device to be calibrated includes: determining the percentage deviation between the first fluorescence data detected in real time and the standard fluorescence data; when the percentage deviation is within the preset percentage deviation range, determining the current working current and working gain as the target working current and target working gain respectively; when the percentage deviation is not within the preset percentage deviation range, continue to adjust the working current and / or working gain until the percentage deviation is within the preset percentage deviation range.

[0085] In an embodiment of the present application, the processor 520 may further be configured to:

[0086] The method further includes: obtaining a first adjustable range of the working gain and a 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.

[0087] In an embodiment of the present application, the processor 520 may further be configured to:

[0088] The method further includes: after determining the target working current and target working gain corresponding to the current channel to be measured, determining the target working current and target working gain corresponding to the next channel to be measured until the calibration of all channels to be measured is completed.

[0089] Through the above solution, by detecting the holes to be measured, the ratio between the actual value and the measured value of the calibration fluorophore is obtained to determine the current adjustment mode or the gain adjustment mode, or by jointly adjusting the current and gain adjustment modes, so as to adjust the working current and working gain of the PCR device to be calibrated. And after determining the target working current and target working gain for all channels to be measured, determine the calibration kingpin of the PCR device to be calibrated. Then, each channel to be measured can perform fluorescence detection based on the adjusted target working current and target working gain. Therefore, the embodiments of the present application can perform linear ratio adjustment of working parameters in different modes for multi-channel PCR devices. Make the adjusted PCR device meet the usage standards, meet the fluorescence performance requirements for instrument production and factory, and improve the accuracy of PCR device detection.

[0090] Figure 6 Schematically shows a schematic diagram of a calibration system for a PCR device according to an embodiment of the present application, as Figure 6As shown in the figure, an embodiment of the present application provides a calibration system for a PCR device, which may include a PCR device 610 to be calibrated and a calibration device 500 for the PCR device. Specifically, the calibration device is the PC side, and the PC side and the PCR device to be calibrated are controlled through communication. The above method for the PCR device is implemented through the calibration software on the PC side. Among them, the calibration software on the PC side includes a first communication module, an in-out bin control module, a first calibration module, and a second calibration module. Specifically, the first communication module can implement communication drive, complete the communication function with the PCR device to be calibrated, and can control the reception of instrument data. The in-out bin control module selects a certain channel for calibration analysis, automatically inserts the calibration fluorophore into the well to be measured through the software, and sets the standard fluorescence data value on the PC side software. The first calibration module collects fluorescence data for the well to be measured, adjusts the LED current and the operational amplifier gain, so that the collected fluorescence data reaches the preset range. The second calibration module uses linear ratio calibration. After the current and gain calibration are completed for the well to be measured, the fluorescence data of all fluxes are collected using the calibrated target working current and target working gain, and reported to the PC side. 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 side software through the network or other communication drives. The calibration analysis module can issue a command through communication on the PC side to make the device start the calibration work. After receiving the command, the instrument to be calibrated starts to work, and reports the fluorescence acquisition data to the PC side, waiting for the PC side to issue 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 side software in real time.

[0091] An embodiment of the present application also provides a machine-readable storage medium, on which instructions are stored, and the instructions are used to make a machine execute the above calibration method for a PCR device.

[0092] In one embodiment, a computer device is provided. The computer device may be a server, and its internal structure diagram may be as Figure 7As shown. The computer device includes a processor A01, a network interface A02, a memory (not shown in the figure), and a database (not shown in the figure) 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 in the figure). 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 the PCR device. The network interface A02 of the computer device is used to communicate with an external terminal through a network connection. When the computer program B02 is executed by the processor A01, it implements a calibration method for the PCR device.

[0093] Those skilled in the art can understand that Figure 7 the structure shown in is only a block diagram of some structures 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 those shown in the figure, or combine certain components, or have different component arrangements.

[0094] Those skilled in the art should understand that the embodiments of the present application can be provided as a method, a system, or a computer program product. Therefore, the present application can take the form of a complete hardware embodiment, a complete software embodiment, or an embodiment combining software and hardware aspects. Moreover, the present application can take the form of a computer program product implemented on one or more computer-usable storage media (including but not limited to disk storage, CD-ROM, optical storage, etc.) containing computer-usable program code.

[0095] The present application is described with reference to the flowcharts and / or block diagrams of methods, devices (systems), and computer program products according to the embodiments of the present application. It should be understood that each flow and / or block in the flowchart and / or block diagram, and the combination of flows and / or blocks in the flowchart and / or block diagram, can be implemented by computer program instructions. These computer program instructions can be provided to the processor of a general-purpose computer, a special-purpose computer, an embedded processor, or other programmable data processing devices to generate a machine, so that the instructions executed by the processor of the computer or other programmable data processing devices generate a device for implementing the functions specified in Figure 1 one process or multiple processes and / or blocks Figure 1 one block or multiple blocks.

[0096] These computer program instructions can also be stored in a computer-readable memory that can direct a computer or other programmable data processing device to work in a particular manner, such that the instructions stored in the computer-readable memory produce a manufacture including an instruction device that implements the functions specified in one process Figure 1 or more processes and / or blocks Figure 1 or more blocks.

[0097] These computer program instructions can also be loaded onto a computer or other programmable data processing device, such that a series of operational steps are performed on the computer or other programmable device to produce a computer-implemented process, whereby the instructions executed on the computer or other programmable device provide steps for implementing the functions specified in one process Figure 1 or more processes and / or blocks Figure 1 or more blocks.

[0098] In a typical configuration, a computing device includes one or more processors (CPUs), an input / output interface, a network interface, and memory.

[0099] The memory may include non-permanent memory in the computer-readable medium, in the form of random access memory (RAM) and / or non-volatile memory, such as read-only memory (ROM) or flash memory (flash RAM). The memory is an example of a computer-readable medium.

[0100] Computer-readable media includes both permanent and non-permanent, removable and non-removable media implemented by any method or technology for storing 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 technologies, compact disc read-only memory (CD-ROM), digital versatile discs (DVD) or other optical storage, magnetic cassettes, magnetic tape 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 media such as modulated data signals and carrier waves.

[0101] It should also be noted that the terms "comprising", "including" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, commodity or device comprising a series of elements not only includes those elements, but also includes other elements not explicitly listed, or elements inherent in such a process, method, commodity or device. Without further limitation, an element defined by the statement "comprising an..." does not exclude the existence of additional identical elements in the process, method, commodity or device comprising the element.

[0102] The above are only embodiments of the present application and are not intended to limit the present application. For those skilled in the art, various changes and modifications can be made to the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall 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 well position to be measured and at least one channel to be measured. The calibration method includes: Controlling each channel to be measured to detect the calibration fluorophore at the well position to be measured to obtain first fluorescence data of the well position to be measured; Obtaining standard fluorescence data of the calibration fluorophore; Determining the ratio between the first fluorescence data and the standard fluorescence data; 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; Adjusting the working current and / or the working gain of the PCR device to be calibrated according to the adjustment mode to determine a target working current and a target working gain corresponding to each channel to be measured, so that each channel to be measured 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, wherein, 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 that the adjustment mode is a current adjustment mode; When the ratio is not within the preset ratio range and the working gain is continuously adjustable, determining that the adjustment mode is a 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 that the adjustment mode is a current adjustment mode and a gain adjustment mode.

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

5. The calibration method for a PCR device according to claim 1, characterized in that, The method further includes: Obtaining a first adjustable range of the working gain and / or a 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 channel to be measured, determining the target working current and the target working gain corresponding to the next channel to be measured until the calibration of all channels to be measured is completed.

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

8. A PCR device, characterized in that, Including: A well position to be measured; At least one channel to be measured; And A calibration device for a PCR device according to claim 8.

9. A calibration system for a PCR device, characterized in that, Including: A PCR device to be calibrated, including a well position to be measured and at least one channel to be measured; And 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.

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