PCR amplification detection system and detection method

Abstract

The invention discloses a PCR (Polymerase Chain Reaction) amplification detection system and a detection method. The PCR amplification detection system comprises a consumable loading module, an optical module and a control module, the consumable loading module is provided with a first loading cavity used for being matched with a first consumable and a second loading cavity used for being matched with a second consumable. The optical module is positioned below the consumable loading module at an interval and is used for performing fluorescence excitation and collection on the consumables in the first loading cavity or the second loading cavity; the control module is electrically connected with the optical module, and the control module is used for controlling the optical module to focus according to the working mode of the PCR amplification detection system, so that the fluorescence excitation and collection of the optical module have the optimal state. According to the invention, compatibility of two consumables in different shapes is realized, more compatibility and convenience are provided for experiments, detection switching of the two loading cavities is realized through focusing processing of the optical module, and relatively high practicability and economical efficiency are achieved.

Description

PCR amplification detection system and detection method Technical Field

[0001] This application belongs to the field of PCR detection technology, specifically relating to a PCR amplification detection system and detection method. Background Technology

[0002] Polymerase chain reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. It can be viewed as a special form of DNA replication outside of a living organism. The most significant characteristic of PCR is its ability to dramatically increase minute amounts of DNA. PCR is a method for the in vitro enzymatic synthesis of specific DNA fragments. It consists of several steps—high-temperature denaturation, low-temperature annealing (renaturation), and optimal-temperature extension—forming a cycle that is repeated cyclically. This allows for rapid amplification of the target DNA, and it is characterized by high specificity, high sensitivity, ease of operation, and time-saving.

[0003] In PCR amplification and detection systems, consumables and reaction reagents are fundamental to PCR experiments. Existing PCR reaction tubes come in various shapes, including round, flat, and multi-tube. However, different shapes of reaction tubes have their own advantages, and different PCR reactions may have different consumable compatibility requirements. However, different PCR instruments can only be matched with consumables of one shape. This limitation of consumable compatibility with a single PCR instrument restricts the scope of PCR instrument use and clearly cannot meet more comprehensive business needs. Summary of the Invention

[0004] The purpose of this application is to provide a PCR amplification detection system and detection method to improve the compatibility of PCR detection equipment with reaction tube consumables of different shapes.

[0005] To achieve the above objectives, this application provides a PCR amplification detection system, the PCR amplification detection system comprising:

[0006] A consumable loading module has a first loading cavity for matching a first consumable and a second loading cavity for matching a second consumable;

[0007] An optical module, spaced below the consumable loading module, is used for fluorescence excitation and collection of consumables within the first or second loading cavity; and

[0008] A control module, electrically connected to the optical module, is used to control the optical module to perform focusing and optimize fluorescence detection according to the working mode of the PCR amplification detection system.

[0009] In an embodiment of this application, the consumable loading module includes:

[0010] A first loading body, wherein the first loading cavity is disposed on the first loading body;

[0011] The second loading body has a second loading cavity disposed on the second loading body and spaced below the first loading body, with the center line of the second loading cavity coinciding with the center line of the first loading cavity;

[0012] A heat-conducting component includes a first heat-conducting element and a second heat-conducting element respectively disposed on the outer peripheral walls of the first loading body and the second loading body, and temperature detection elements are respectively provided on the first heat-conducting element and the second heat-conducting element;

[0013] A temperature control component is electrically connected to the temperature detection element. The temperature control component is used to receive the temperature signal sent by the temperature detection element and to regulate the temperature of the second consumable or the first consumable according to the temperature signal.

[0014] In embodiments of this application, the temperature control component includes:

[0015] The first temperature control is located on the side of the first heat-conducting component away from the outer peripheral wall of the first loading body. The first temperature control is used to regulate the temperature of the first consumable in the first loading cavity.

[0016] The second temperature control is located on the inner peripheral wall of the second loading cavity and is used to regulate the temperature of the second consumable within the second loading cavity.

[0017] In an embodiment of this application, the temperature control component further includes a third temperature control device, which is disposed on the side of the second heat-conducting element away from the outer peripheral wall of the second loading body. The third temperature control device cooperates with the second temperature control device to regulate the temperature of the second consumable in the second loading cavity.

[0018] In an embodiment of this application, the second loading body includes two loading sections joined together, the two loading sections being arranged opposite each other to form the second loading cavity, and the consumable loading module further includes:

[0019] A connecting rod is connected to each of the two loading parts;

[0020] A driving component is linearly driven to the connecting rods, and the driving component is used to drive the two connecting rods to move linearly, so as to move the two loading parts closer to or further away from each other.

[0021] In embodiments of this application, the optical module includes an integrated component:

[0022] The excitation assembly includes a light source, a collimating lens, and a first filter arranged sequentially along the optical path transmission direction;

[0023] The acquisition component includes a second filter, a focusing lens, and a photodetector arranged sequentially along the optical path transmission direction;

[0024] A zoom lens assembly, wherein the emitting end of the zoom lens assembly is aligned with the centerline of the consumable and excites and collects fluorescence from the sample within the consumable;

[0025] The dichroic mirror transmits the fluorescence excited by the excitation component to the zoom lens component, and the dichroic mirror can also transmit the fluorescence emitted by the zoom lens component to the acquisition component.

[0026] In embodiments of this application, the zoom lens assembly includes two lenses arranged vertically at a distance from each other, and the focal length of the zoom lens assembly needs to meet the following conditions:

[0027]

[0028] Where f is the focal length of the zoom lens assembly, f1 is the focal length of the first lens, f2 is the focal length of the second lens, and d is the distance between the two lenses.

[0029] A second aspect of this application provides a PCR amplification detection method, applied to the PCR amplification detection system described above, the PCR amplification detection method comprising the following steps:

[0030] Select the desired work mode;

[0031] Load the consumables into the corresponding loading chamber;

[0032] The optical module is focused to its optimal state according to the required operating mode.

[0033] The temperature control component is activated to individually regulate the temperature of the consumables in the loading cavity, so that either the first or second consumable is kept within the preset temperature range.

[0034] The optical module is activated to excite and collect fluorescence from the consumables within the loading cavity.

[0035] In an embodiment of this application, the step of focusing the optical module to the optimal state according to the required operating mode includes:

[0036] When the required working mode is fixed focus mode, control the zoom lens assembly to optically fix the focus to the preset position;

[0037] When the required operating mode is the optimization mode, the zoom lens assembly is controlled to find the optimal optical focusing position, and the zoom lens assembly is adjusted to focus according to the optimal optical focusing position.

[0038] In the embodiments of this application, the step of controlling the zoom lens assembly to find the optimal optical focusing position and adjusting the zoom lens assembly to focus according to the optimal optical focusing position when the required working mode is the optimization mode includes:

[0039] Obtain the preset focal length position of the zoom lens assembly;

[0040] Multiple focus points are scanned near the fixed-focus preset position, and the fluorescence acquisition signal value of each focus point is obtained;

[0041] The optimal optical focusing position is selected from among multiple focusing points that has the largest fluorescence acquisition signal value.

[0042] In embodiments of this application, the step of activating the temperature control component to individually regulate the temperature of the consumables within the loading cavity, so that either the first or second consumable is maintained within a preset temperature range, further includes:

[0043] During the fluorescence excitation and collection process of the second consumable, the temperature of the second consumable is acquired in real time and the second temperature control is activated.

[0044] When the temperature of the second consumable changes rapidly, the second temperature control and the third temperature control are activated simultaneously so that the second temperature control and the third temperature control work together.

[0045] Through the above technical solutions, the PCR amplification and detection system provided by the embodiments of the present invention has the following beneficial effects:

[0046] This application enables the consumable loading device to accommodate different types of reaction tubes by forming a first loading cavity for matching a first consumable and a second loading cavity for matching a second consumable on the consumable loading module. Before experimental operation, the shape of the consumable is determined, and the corresponding consumable is installed in the first or second loading cavity. The control module controls the optical module to perform focusing optimization detection, ensuring optimal fluorescence excitation and acquisition. Then, the optical module is activated to excite and acquire fluorescence from the consumable. This application not only achieves compatibility with two different shaped consumables, providing greater compatibility and convenience for experiments, but also allows for independent control and detection switching between the two loading cavities through the focusing process of the optical module, demonstrating strong practicality and economy.

[0047] Other features and advantages of the embodiments of this application will be described in detail in the following detailed description section. Attached Figure Description

[0048] The accompanying drawings are provided to further illustrate the embodiments of this application and form part of the specification. They are used together with the following detailed description to explain the embodiments of this application, but do not constitute a limitation on the embodiments of this application. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without any inventive effort. In the drawings:

[0049] Figure 1 is a schematic diagram of the PCR amplification detection system according to this application;

[0050] Figure 2 is a schematic diagram of the PCR amplification detection system according to this application when detecting flat tube consumables;

[0051] Figure 3 is a schematic diagram of the PCR amplification detection system according to this application when detecting round tube consumables;

[0052] Figure 4 is a schematic diagram of the optical module in the PCR amplification detection system according to this application;

[0053] Figure 5 is a schematic diagram of the zoom lens assembly in the optical module of this application;

[0054] Figure 6 is a flowchart illustrating the PCR amplification detection method of this application.

[0055] Explanation of reference numerals in the attached figures

[0056] 11 First loading body 63 First filter

[0057] 12 Second loading body 64 dichroic mirror

[0058] 13 First loading cavity 65 Collimating lens

[0059] 14 Second loading cavity 66 Second filter

[0060] 21 First heat-conducting component 67 Photodetector

[0061] 22 Second heat-conducting component 68 Zoom lens assembly

[0062] 31 First temperature control unit 691 First lens

[0063] 32 Second temperature control 692 Second lens

[0064] 33 Third temperature control unit 80 Connecting rod

[0065] 50 mounting holes, 100 consumable loading module

[0066] 61 Light Source 200 Optical Module

[0067] 62 Focusing Lens Detailed Implementation

[0068] The specific embodiments of this application will be described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are for illustration and explanation only and are not intended to limit this application.

[0069] The PCR amplification detection system and detection method according to this application are described below with reference to the accompanying drawings.

[0070] As shown in Figure 1, in an embodiment of this application, a PCR amplification detection system is proposed. The PCR amplification detection system includes a consumable loading module 100, an optical module 200, and a control module. The consumable loading module 100 has a first loading cavity 13 for matching a first consumable and a second loading cavity 14 for matching a second consumable. The optical module 200 is located below the consumable loading module 100 and is used to excite and collect fluorescence from the consumables in the first loading cavity 13 or the second loading cavity 14. The control module is electrically connected to the optical module 200 and is used to control the optical module 200 to focus according to the working mode of the PCR amplification detection system so that the fluorescence excitation and collection of the optical module 200 are in the optimal state.

[0071] Before the experiment, the shape of the reaction tube is determined, and the corresponding reaction tube is installed in the first loading cavity 13 or the second loading cavity 14. The optical module 200 is controlled by the control module to perform focusing optimization detection so that the fluorescence excitation and acquisition of the optical module 200 are in the best state. Then, the optical module 200 is turned on to excite and acquire fluorescence of the consumables. This application not only achieves compatibility with two different shaped consumables, providing more compatibility and convenience for the experiment, but also allows the two loading cavities to be independently controlled and switched by adjusting the focusing method of the optical module 200, which has strong practicality and economy.

[0072] It should be noted that the shapes of the first and second consumables in this application can be the same or different. The number of consumables can also be multiple; this application uses two as an example for explanation. When the first and second consumables have the same shape, both can be round tubes, flat tubes, or square tubes, etc.; when the shapes of the first and second consumables are different, for ease of understanding, the following explanation uses a round tube as the first consumable and a flat tube as the second consumable.

[0073] In the embodiments of this application, the consumable loading module 100 includes a first loading body 11, a second loading body 12, a heat-conducting component, and a temperature control component; a first loading cavity 13 is disposed on the first loading body 11; a second loading cavity 14 is disposed on the second loading body 12 and spaced below the first loading body 11, the center line of the second loading cavity 14 coincides with the center line of the first loading cavity 13; the heat-conducting component includes a first heat-conducting element 21 and a second heat-conducting element 22 respectively disposed on the outer peripheral walls of the first loading body 11 and the second loading body 12, the first heat-conducting element 21 and the second heat-conducting element 22 respectively provided with temperature detection elements for detecting the corresponding loading cavities, the temperature detection elements are preferably temperature sensors, and mounting holes 50 for mounting temperature sensors are opened on the first heat-conducting element 21 and the second heat-conducting element 22; the temperature control component is electrically connected to the temperature detection elements, and the temperature control component is used to receive the temperature signal sent by the temperature detection elements and to independently regulate the temperature of the flat tube consumables and the round tube consumables according to the temperature signal.

[0074] In this embodiment, the first loading body 11 is fixed in position by an external connector, and the lower second loading body 12 is connected to another component. The two loading bodies are independently set and do not interfere with each other. Both the first heat-conducting element 21 and the second heat-conducting element 22 can be metal blocks with good thermal conductivity, such as copper or some alloy components, to achieve good heat conduction. The temperature control component can be a semiconductor cooler (TEC), etc. Temperature sensors are provided in both the first heat-conducting element 21 and the second heat-conducting element 22 to provide real-time feedback on the temperature information of the corresponding consumables and adjust the temperature control component. Precise control of the temperature of the consumable loading chamber is achieved through temperature program settings, providing the temperature conditions required for the PCR experimental reaction.

[0075] In embodiments of this application, the temperature control component includes:

[0076] The first temperature control 31 is located on the side of the first heat-conducting component 21 away from the outer peripheral wall of the first loading body 11. The first temperature control 31 is used to regulate the temperature of the circular tube consumable in the first loading cavity 13.

[0077] The second temperature control unit 32 is located on the inner peripheral wall of the second loading cavity 14 and is used to regulate the temperature of the flat tube consumables inside the second loading cavity 14.

[0078] In actual operation, the first loading chamber 13 and the second loading chamber 14 have independent temperature control modes. The upper round tube consumable is regulated by the first temperature control 31 on the periphery of the first loading body 11; the lower flat tube consumable is regulated by the second temperature control 32, thus achieving independent temperature control for the upper and lower parts. Specifically, temperature sensors acquire the temperatures of the first loading chamber 13 and the second loading chamber 14 respectively, and adjust the temperature of the corresponding loading chamber through the corresponding temperature control, thereby ensuring that the temperature of the consumables in the loading chamber reaches a suitable reaction range.

[0079] In the embodiments of this application, the outer periphery of the loading body is connected to a corresponding heat sink, such as a fan, to meet the rapid heat dissipation requirements during temperature control.

[0080] Furthermore, there are two sets of second temperature control units 32, which are arranged opposite each other on the inner peripheral wall of the second loading cavity 14. The second temperature control unit 32 has a block-shaped structure and its area is smaller than that of the first temperature control unit 31. During experiments, the second temperature control unit 32 can fit tightly against the flat tube consumable, providing faster and more efficient temperature regulation and improving PCR experimental efficiency.

[0081] In the embodiments of this application, the temperature control component further includes a third temperature control device 33, which is disposed on the side of the second heat-conducting element 22 away from the outer peripheral wall of the second loading body 12. The third temperature control device 33 cooperates with the second temperature control device 32 to regulate the temperature of the flat tube consumable in the second loading cavity 14.

[0082] A third temperature control unit 33 is also provided on the second loading body 12. The third temperature control unit 33 is located on the periphery of the second heat-conducting element 22. The third temperature control unit 33 works in conjunction with the second temperature control unit 32 to achieve rapid temperature change of the flat tube consumable. Of course, in some cases, if a more gentle temperature change condition is required, or for cost considerations, one or both of the third temperature control units 33 can be removed, allowing the flat tube consumable surface to directly contact the second heat-conducting element 22, and the temperature can be regulated by the second temperature control unit 32 on the inner side wall.

[0083] In the embodiments of this application, the second loading body 12 includes two loading parts spliced ​​together. The two loading parts are arranged opposite to each other and form a second loading cavity 14. The consumable loading module 100 also includes a connecting rod 80 and a driving member. A connecting rod 80 is connected to each of the two loading parts. The driving member is linearly driven to the connecting rod 80. The driving member is used to drive the two connecting rods 80 to move linearly, so as to drive the two loading parts to move closer or further away from each other.

[0084] In this application, since the center lines of the first loading cavity 13 and the second loading cavity 14 coincide, and the second loading cavity 14 is located below the first loading cavity 13, and since the detection system can only detect one type of consumable at a time, when detecting round tube consumables, the light source 61 enters the first loading cavity 13 through the second loading cavity 14. To improve fluorescence excitation and collection efficiency, the driving component drives the two connecting rods 80 to move in opposite directions, so that the two loading parts move away from each other, thereby increasing the inner diameter of the second loading cavity 14 and avoiding light obstruction. When detecting flat tube consumables, the upper first loading body 11 is fixed in position, and the second loading cavity 14 formed in the lower second loading body 12 clamps the flat tube consumable to achieve detection of the flat tube consumable.

[0085] In addition, the first loading body 11 also includes two loading parts assembled together. The two loading parts are arranged opposite to each other, and the opposite surfaces of the two loading parts form concave surfaces. The two concave surfaces cooperate with each other to form a first loading cavity 13 that matches the circular tube consumable. A first heat-conducting element 21 is disposed on the outer periphery of the two loading parts, and a first temperature control element 31 is installed on the outer peripheral wall of the first heat-conducting element 21.

[0086] As shown in Figure 4, the optical module 200 includes an integrated unit:

[0087] The excitation assembly includes a light source 61, a collimating lens 65, and a first filter 63 arranged sequentially along the optical path transmission direction.

[0088] The acquisition component includes a second filter 66, a focusing lens 62, and a photodetector 67 arranged sequentially along the optical path transmission direction.

[0089] The zoom lens assembly 68 has its emitting end aligned with the center line of the consumable and emits fluorescence to excite the consumable's emission assembly.

[0090] The dichroic mirror 64 transmits the fluorescence excited by the excitation component to the zoom lens assembly 68, and the dichroic mirror 64 can also transmit the fluorescence emitted by the zoom lens assembly 68 to the acquisition component.

[0091] Preferably, the zoom lens assembly 68 can be a liquid lens, and the lens shape can be changed by adjusting the parameters of the liquid lens, thereby changing the lens focal length. It can also be achieved by combining multiple lenses.

[0092] This application integrates an optical module 200 into one unit. In terms of fluorescence signal excitation and acquisition, the focal position of the optical module 200 is changed by adjusting the zoom lens assembly 68, thereby achieving fluorescence excitation and acquisition at different loading cavity positions, reducing the size and improving the ease of instrument operation.

[0093] The optical module 200 is shown in Figure 4. The specific optical path is as follows: excitation light emitted from light source 61 (which can be an LED, laser, etc.) is collimated by collimating lens 65, then filtered by first filter 63, selecting a specific wavelength of light to pass through. This light then passes through dichroic mirror 64, is focused by zoom lens assembly 68, and enters the loading cavity to irradiate the sample. Since the sample contains fluorescent substances, fluorescence is excited. The fluorescence signal generated by the fluorescent substances is collimated by the same zoom lens assembly 68, reflected by dichroic mirror 64, filtered by second filter 66, selecting a specific wavelength of fluorescence to pass through, and then focused by focusing lens 62 onto photodetector 67 to detect the fluorescence signal. Figure 4 only illustrates the excitation and acquisition of fluorescence signals from one channel. In reality, the optical module 200 can be a combination module with multiple color channels of different wavelengths, such as red, yellow, green, and blue color combinations. Through folded or stepped optical paths, multiple channels can be combined to achieve multiplex PCR experimental detection.

[0094] Preferably, as shown in FIG5, the zoom lens assembly 68 includes two lenses arranged vertically at a distance, and the focal length of the zoom lens assembly 68 needs to meet the following conditions:

[0095]

[0096] Where f is the focal length of the zoom lens assembly 68, f1 is the focal length of the first lens 691, f2 is the focal length of the second lens 692, and d is the distance between the two lenses.

[0097] In this embodiment, the focal length of the zoom lens assembly 68 can be adjusted by adjusting the distance between the two lenses in the zoom lens assembly 68, thereby enabling the switching of fluorescence excitation acquisition between the two loading cavities.

[0098] As shown in Figure 6, a second aspect of this application provides a PCR amplification detection method, applied to the PCR amplification detection system described above. The PCR amplification detection method includes the following steps:

[0099] Select the desired work mode;

[0100] Load the consumables into the corresponding loading chamber;

[0101] The optical module 200 is focused to the optimal state according to the required working mode;

[0102] The optical module 200 is activated to excite and collect fluorescence from the consumables inside the loading cavity.

[0103] The detection system of this application can only be used in the case of single tube operation, that is, only one loading chamber 13 or the second loading chamber 14 needs to be selected for use, while the other loading chamber is left empty.

[0104] The operating modes are either fixed-focus mode or optimized mode. In fixed-focus mode, focusing is only required by controlling the optical module 200 according to the fixed focus point. In optimized mode, the optical module 200 needs to be adjusted to find the optimal focus point for focusing. This application uses the zoom function of the optical module 200 to achieve independent control and detection switching of the two loading chamber PCR experiments. This not only achieves compatibility with two different shaped consumables, providing greater compatibility and convenience for experiments, but also greatly reduces the size of the instrument by achieving detection switching through simple zoom, making it highly practical and convenient.

[0105] In the embodiments of this application, the step of controlling the optical module 200 to focus according to the required working mode includes:

[0106] When the required working mode is fixed focus mode, control the zoom lens assembly 68 to optically fix focus to the preset position;

[0107] When the required operating mode is the optimization mode, the zoom lens assembly 68 is controlled to find the optimal optical focusing position, and the zoom lens assembly 68 is adjusted to focus according to the optimal optical focusing position.

[0108] In the fixed-focus operation, which is the default mode, the focus points of the zoom lens assembly 68 in the two loading cavities are preset through calculation and matching, for example, F1 and F2. According to the shape of the consumables, they are loaded into the corresponding loading cavity structures, and then the zoom is switched. For the lower second loading cavity 14, the focus is fixed at F1, and for the upper first loading cavity 13, the focus is fixed at F2. Then, the PCR cycle experiment is started, and the temperature-controlled PCR amplification and fluorescence excitation and acquisition are cycled to achieve real-time monitoring of the PCR reaction.

[0109] In fixed-focus mode, as shown in Figures 2 and 3, by adjusting the lens focal length, the focus is fixed at the center point F1 of the lower consumable cavity, enabling the optical module 200 to excite and collect fluorescence from the lower consumable cavity. As shown in Figure 6, by adjusting the lens focal length, the focus is fixed at the center point F2 of the upper consumable cavity, enabling the optical module 200 to excite and collect fluorescence from the upper consumable cavity.

[0110] Furthermore, in a preferred embodiment, in order to improve the efficiency of fluorescence excitation and collection of the sample in the first loading cavity, the two loading parts on both sides of the second loading cavity 14 can be adjusted to move away from each other, thereby increasing the inner diameter of the second loading cavity 14 and avoiding light blockage.

[0111] In another embodiment, the fluorescence excitation and acquisition signal intensity can be increased by adjusting the light emission intensity of the excitation component, thereby improving the fluorescence detection sensitivity.

[0112] In optimization mode, the optimal focus point is found nearby based on the preset top focus point. For example, the optimal focus point F1' is found near F1, and the optimal focus point F2' is found near F2. The zoom is switched according to the found optimal focus point. For the second loading cavity 14, the focus is fixed at F1', and for the first loading cavity 13, the focus is fixed at F2'.

[0113] In the embodiments of this application, the step of controlling the zoom lens assembly 68 to find the optimal optical focusing position and adjusting the zoom lens assembly 68 to focus according to the optimal optical focusing position when the required working mode is the optimization mode includes:

[0114] Obtain the fixed-focus preset position of the zoom lens assembly 68;

[0115] Multiple focus points are scanned near the fixed-focus preset position, and the fluorescence acquisition signal value of each focus point is obtained;

[0116] The optimal optical focusing position is selected from among multiple focusing points that has the largest fluorescence acquisition signal value.

[0117] In optimization mode, the optimal optical focusing position for the experiment is found through program optimization of the control module. Specifically, after placing the specific consumables and reagents and before the experiment begins, the fluorescence excitation and acquisition signal is fixed. The zoom lens assembly 68 is adjusted to scan a series of focusing points near the fixed focus point, for example, scanning 10 equally spaced focusing points. The focusing point with the largest fluorescence acquisition signal value is determined as the optimal focusing position for this experiment. Optical focusing is then used to start the PCR cycle experiment, cyclically performing temperature-controlled PCR amplification and fluorescence excitation and acquisition, achieving real-time monitoring of the PCR amplification experiment. Using optimization mode can greatly improve the fluorescence excitation and acquisition efficiency, effectively calibrate and compensate for assembly deviations of experimental modules and consumables, and improve the consistency of experimental results.

[0118] In the embodiments of this application, the PCR amplification detection method further includes the following steps:

[0119] Before fluorescence excitation and collection of consumables, the temperature control component is turned on to individually regulate the temperature of the consumables in the loading cavity, so that both round tube consumables and flat tube consumables are kept within the preset temperature range.

[0120] Since the detection system can only operate on a single tube, meaning that round tube consumables and flat tubes cannot be tested simultaneously but are tested separately; during the fluorescence excitation and collection of consumables, only a temperature control component is needed to individually control the temperature of the consumables in the corresponding loading chamber, so that the round tube consumables or flat tube consumables are within a preset temperature range when working alone.

[0121] In the embodiments of this application, the step of activating the temperature control component and individually regulating the temperature of the consumables in the loading cavity before fluorescence excitation and collection of the consumables, so that both the round tube consumables and the flat tube consumables are kept within a preset temperature range, further includes:

[0122] Before performing fluorescence excitation and acquisition on the flat tube consumable, the second temperature control 32 is turned on and the temperature of the flat tube consumable is acquired in real time.

[0123] When rapid temperature changes are required for the flat tube consumable, the second temperature control 32 and the third temperature control 33 are activated simultaneously so that they work together.

[0124] When performing fluorescence excitation and acquisition on flat tube consumables, it is necessary to monitor the temperature of the consumables in real time. When the consumables are within a relatively stable temperature range, only the second temperature control 32 needs to be activated to regulate the temperature. If the temperature of the consumables needs to be rapidly increased or decreased during the experiment, the external third temperature control 33 can be activated. The third temperature control 33 and the second temperature control 32 are used in conjunction to meet the temperature regulation needs of the flat tube consumables, providing faster and more efficient temperature changes and improving the efficiency of PCR experiments. When performing fluorescence excitation and acquisition on round tube consumables, only the first temperature control 31 needs to be activated to regulate the temperature.

[0125] In the description of this application, it should be understood that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0126] In this application, unless otherwise expressly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection, an electrical connection, or a connection that allows communication between components; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components, unless otherwise expressly limited. Those skilled in the art can understand the specific meaning of the above terms in this application according to the specific circumstances.

[0127] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0128] Although embodiments of this application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting this application. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of this application.

Claims

1. A PCR amplification detection system, characterized in that, The PCR amplification detection system includes: a consumable loading module (100) having a first loading cavity (13) for matching a first consumable and a second loading cavity (14) for matching a second consumable; an optical module (200) spaced below the consumable loading module (100) and used for fluorescence excitation and acquisition of consumables in the first loading cavity (13) or the second loading cavity (14); and a control module electrically connected to the optical module (200), the control module being used to control the optical module (200) to perform focusing optimization fluorescence detection according to the working mode of the PCR amplification detection system.

2. The PCR amplification detection system according to claim 1, characterized in that, The consumable loading module (100) includes: a first loading body (11), wherein the first loading cavity (13) is opened on the first loading body (11); a second loading body (12), wherein the second loading cavity (14) is opened on the second loading body (12) and is spaced below the first loading body (11), and the center line of the second loading cavity (14) coincides with the center line of the first loading cavity (13); a heat-conducting component, including a first heat-conducting element (21) and a second heat-conducting element (22) respectively disposed on the outer peripheral walls of the first loading body (11) and the second loading body (12), wherein the first heat-conducting element (21) and the second heat-conducting element (22) are respectively provided with temperature detection elements; and a temperature control component, electrically connected to the temperature detection elements, wherein the temperature control component is used to receive the temperature signal sent by the temperature detection elements and to regulate the temperature of the second consumable or the first consumable according to the temperature signal.

3. The PCR amplification detection system according to claim 2, characterized in that, The temperature control component includes: a first temperature control (31), disposed on the side of the first heat-conducting element (21) away from the outer peripheral wall of the first loading body (11), the first temperature control (31) being used to regulate the temperature of the first consumable in the first loading cavity (13); and a second temperature control (32), the inner peripheral wall of the second loading cavity (14) being used to regulate the temperature of the second consumable in the second loading cavity (14).

4. The PCR amplification detection system according to claim 3, characterized in that, The temperature control component also includes a third temperature control unit (33), which is located on the side of the second heat-conducting element (22) away from the outer peripheral wall of the second loading body (12). The third temperature control unit (33) cooperates with the second temperature control unit (32) to regulate the temperature of the second consumable in the second loading cavity (14).

5. The PCR amplification detection system according to claim 2, characterized in that, The second loading body (12) includes two loading parts spliced ​​together. The two loading parts are arranged opposite to each other and form the second loading cavity (14). The consumable loading module (100) also includes: a connecting rod (80), one of which is connected to each of the two loading parts; and a driving member, which is linearly driven connected to the connecting rod (80). The driving member is used to drive the two connecting rods (80) to move linearly, so as to drive the two loading parts to move closer or further away from each other.

6. The PCR amplification detection system according to any one of claims 1 to 5, characterized in that, The optical module (200) includes: an excitation component, comprising a light source (61), a collimating lens (65), and a first filter (63) arranged sequentially along the optical path transmission direction; a collection component, comprising a second filter (66), a focusing lens (62), and a photodetector (67) arranged sequentially along the optical path transmission direction; a zoom lens component (68), the emitting end of which is aligned with the center line of the consumable and excites and collects fluorescence from the sample within the consumable; and a dichroic mirror (64), through which the fluorescence excited by the excitation component is transmitted to the zoom lens component (68), and the dichroic mirror (64) can transmit the fluorescence emitted by the zoom lens component (68) to the collection component.

7. The PCR amplification detection system according to claim 6, characterized in that, The zoom lens assembly (68) includes two lenses arranged vertically at a distance from each other. The focal length of the zoom lens assembly (68) needs to meet the following conditions: Where f is the focal length of the zoom lens assembly (68), f1 is the focal length of the first lens (691), f2 is the focal length of the second lens (692), and d is the distance between the two lenses.

8. A PCR amplification detection method, characterized in that, The PCR amplification detection method, applied to any one of claims 1 to 7, comprises the following steps: selecting the desired working mode; loading consumables into the corresponding loading chamber; focusing the optical module (200) to the optimal state according to the desired working mode; activating the temperature control component to individually regulate the temperature of the consumables in the loading chamber, so that either the first or second consumable is maintained within a preset temperature range; and activating the optical module (200) to excite and collect fluorescence from the consumables in the loading chamber.

9. The PCR amplification and detection method according to claim 8, characterized in that, The step of focusing the optical module (200) to the optimal state according to the required working mode includes: when the required working mode is fixed focus mode, controlling the zoom lens assembly (68) to optically fix focus to a preset position; when the required working mode is optimization mode, controlling the zoom lens assembly (68) to find the optimal optical focusing position, and adjusting the zoom lens assembly (68) to focus according to the optimal optical focusing position.

10. The PCR amplification and detection method according to claim 9, characterized in that, When the required working mode is optimization mode, the steps of controlling the zoom lens assembly (68) to find the optimal optical focusing position and adjusting the zoom lens assembly (68) to focus according to the optimal optical focusing position include: obtaining the fixed focus preset position of the zoom lens assembly (68); scanning multiple focus points near the fixed focus preset position and obtaining the fluorescence acquisition signal value of each focus point; selecting the focus point position with the largest fluorescence acquisition signal value among the multiple focus points as the optimal optical focusing position.

11. The PCR amplification and detection method according to claim 8, characterized in that, The step of activating the temperature control component to individually regulate the temperature of the consumables in the loading cavity so that the first consumable or the second consumable is kept within a preset temperature range further includes: before fluorescence excitation and collection of the second consumable, acquiring the temperature of the second consumable in real time and activating the second temperature control (32); when a rapid temperature change of the second consumable is required, activating the second temperature control (32) and the third temperature control (33) simultaneously so that the second temperature control (32) and the third temperature control (33) work together.

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