Centrifugal nucleic acid detection equipment and detection system based on image analysis

Abstract

The utility model provides centrifugal nucleic acid detection equipment and a detection system based on image analysis, and the equipment comprises a rack, and the following structures arranged on the rack: a rotating module; a temperature control module; an irradiation module; an imaging module; and a data processing module. According to the equipment provided by the utility model, only the imaging module is needed to shoot the area, where the micro-fluidic chip is arranged, of the rotating module to obtain the image data, and the data processing module is used for processing the image data to obtain the nucleic acid detection result, so that compared with the prior art, the equipment provided by the invention is lower in cost and more convenient to use. And meanwhile, the detection efficiency is also greatly improved.

Description

Technical Field

[0001] The present application relates to the technical field of molecular in vitro diagnosis, and in particular to a centrifugal nucleic acid detection device and detection system based on image analysis. Background Art

[0002] Point-of-care testing, as a fast and accurate detection technology, is used in large-scale screening and has also been recognized by the testing market. It is widely used in non-medical fields such as the detection of viruses and bacteria in various forms of samples such as respiratory tract and blood, as well as sample species identification. In point-of-care testing technology, antigen-based colloidal gold paper strip tests are widely used. However, antigen detection has low sensitivity and is prone to missed detection. At the same time, colloidal gold paper strips are exposed to the testing environment and are easily affected by the environment and sample conditions, resulting in false positives.

[0003] In comparison, nucleic acid molecule-based detection technologies, especially those based on polymerase chain reaction (PCR) technology, are used as standard detection methods in nucleic acid detection and other fields due to their high sensitivity and high specificity. However, PCR detection technology has relatively high requirements for equipment and cannot meet the needs of instant detection in simple or non-professional scenarios. Isothermal amplification detection technology does not require precise temperature control, has low requirements for equipment, has better sensitivity and specificity than antigen detection, and its detection accuracy is close to that of PCR detection. Therefore, it is the most commonly used technology in current instant detection products.

[0004] In the molecular detection technology equipment currently on the market and reported in the literature, especially the instruments using isothermal amplification detection technology, the signal reporting method used is mainly fluorescence or visible light colorimetry, and the changes of these fluorescence or visible light signals are converted into electrical signals through the built-in photoelectric sensor and displayed on the instrument reporting end. Such a reporting method has certain requirements for the performance of the photoelectric sensor, and the detection of multiple samples and multiple targets will increase the cost of the photoelectric sensor or related components, making it difficult to apply to instant detection scenarios with low-cost requirements. Utility Model Content

[0005] In order to solve the problems in the prior art, the present application provides a centrifugal nucleic acid detection device and detection system based on image analysis. The technical solution of the present application is as follows:

[0006] A centrifugal nucleic acid detection device based on image analysis, comprising:

[0007] A frame, and the following structures arranged on the frame:

[0008] A rotating module, wherein the rotating module can carry the microfluidic chip and drive the microfluidic chip to rotate;

[0009] A temperature control module, which can control the temperature of the microfluidic chip carried by the rotating module so that the sample inside the microfluidic chip can undergo amplification reaction and gene shearing;

[0010] An irradiation module, which can emit light to stimulate the samples undergoing amplification reaction and gene shearing inside the microfluidic chip to emit fluorescence;

[0011] an imaging module, which can photograph the area on the rotating module where the microfluidic chip is arranged to obtain image data when the sample after amplification reaction and gene shearing inside the microfluidic chip emits fluorescence; and,

[0012] A data processing module is electrically connected to the imaging module to receive the image data and process the image data to obtain a nucleic acid detection result.

[0013] Furthermore, the rotation module includes: a motor, which is directly or indirectly fixed to the frame; and a tray, which is connected to the rotating shaft of the motor so that the rotating shaft of the motor can drive the tray to rotate, and the tray can carry and fix more than one microfluidic chip.

[0014] Furthermore, the rotating module further comprises: a tray fixing seat, the tray fixing seat is fixedly connected to the rotating shaft of the motor, and the tray fixing seat can be fixedly connected to the tray.

[0015] Furthermore, the rotation module further comprises: a chip pressing cover, and the chip pressing cover can press the microfluidic chip onto the tray.

[0016] Furthermore, the temperature control module includes: a temperature control tray assembly, which is located on one side of the tray; a temperature control component, which is arranged on the side of the temperature control tray assembly facing the tray, and the temperature control component can control the temperature of the microfluidic chip.

[0017] Furthermore, the temperature control component is a PI heating film.

[0018] Furthermore, the temperature control module further includes: a motion component, and the motion component can drive the temperature control tray component to move closer to or away from the tray.

[0019] Furthermore, the motion assembly includes: a motion unit, which is directly or indirectly fixedly connected to the frame; and a motion bracket, through which the motion unit can drive the temperature-controlled tray assembly to move closer to or away from the tray.

[0020] Furthermore, the temperature-controlled tray assembly includes: a first temperature-controlled tray unit; a second temperature-controlled tray unit, the second temperature-controlled tray unit is arranged in parallel with the first temperature-controlled tray unit, and the second temperature-controlled tray unit and the tray are located on different sides of the first temperature-controlled tray unit; an elastic member, the elastic member is arranged between the first temperature-controlled tray unit and the second temperature-controlled tray unit; wherein the motion assembly can drive the second temperature-controlled tray unit to approach or move away from the tray.

[0021] Furthermore, the temperature control module also includes: a temperature detection component, and the temperature detection component can detect the temperature of the microfluidic chip.

[0022] Furthermore, the irradiation module includes: a lampshade, which is arranged on the outside of the tray; and a plurality of light sources, which are arranged on the inside of the lampshade, and the light emitted by the light sources irradiates the microfluidic chip carried on the rotating module from the side.

[0023] Furthermore, the imaging module includes: an image sensor; a lens assembly; and a filter, which can filter out the light emitted by the irradiation module and retain the fluorescence emitted by the amplification reaction inside the microfluidic chip and the sample after gene shearing.

[0024] Furthermore, the data processing module obtains the detection result based on the YOLO algorithm.

[0025] Furthermore, a tray mark is provided on the tray; and the data processing module is capable of acquiring the position information of the microfluidic chip at least according to the information of the tray mark in the image data.

[0026] Furthermore, a chip mark is provided on the microfluidic chip; and the data processing module is at least capable of distinguishing different microfluidic chips according to the information of the chip mark in the image data.

[0027] Furthermore, the data processing module is also electrically connected to the rotation module to control the rotation module to drive the microfluidic chip to rotate; the data processing module is also electrically connected to the temperature control module to control the temperature control module to perform temperature control; and / or, the data processing module is also electrically connected to the irradiation module to control the irradiation module to emit light to stimulate the amplification reaction and the sample after gene shearing to emit fluorescence.

[0028] Further, it includes: any of the above-mentioned centrifugal nucleic acid detection devices based on image analysis; and a microfluidic chip.

[0029] The centrifugal nucleic acid detection device based on image analysis provided by the present application does not require a photoelectric sensor. It only requires a more mature, existing and lower-cost imaging module to shoot the area where the microfluidic chip is set on the rotating module to obtain image data, and the data processing module processes the image data to obtain the nucleic acid detection result. In particular, when multiple detection cavities are set on the microfluidic chip and / or multiple microfluidic chips are set on the device at the same time, the imaging module can shoot the entire area where the microfluidic chip is set on the rotating module once to obtain image data, and the data processing module processes the image data once to obtain the detection results of all detection cavities in all microfluidic chips. Therefore, compared with the prior art, the above-mentioned equipment of the present application is lower in cost and also greatly improves the detection efficiency.

[0030] The above description is only an overview of the technical solution of the present application. In order to make the technical means of the present application clearer and to enable those skilled in the art to implement it according to the contents of the specification, and to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are exemplified below. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 : A side view of a centrifugal nucleic acid detection device based on image analysis in one embodiment;

[0032] Figure 2 : A left view of a centrifugal nucleic acid detection device based on image analysis in one embodiment;

[0033] Figure 3 : Figure 2 Schematic diagram of the middle DD section;

[0034] Figure 4 : A schematic diagram of data processing module connection in one embodiment;

[0035] Figure 5 : Schematic diagram of the structure of a microfluidic chip in one embodiment.

[0036] Description of reference numerals:

[0037] A. Centrifugal nucleic acid detection equipment based on image analysis;

[0038] 110a, bottom plate; 120a, vertical plate;

[0039] 210a, motor; 220a, tray; 230a, tray fixing seat; 240a, chip cover; 250a, nut; 260a, motor fixing seat;

[0040] 310a, temperature control tray assembly; 311a, first temperature control tray unit; 312a, second temperature control tray unit; 313a, elastic member; 320a, temperature control assembly; 330a, motion assembly; 331a, fixed bracket; 332a, motion unit; 333a, motion bracket; 340a, temperature detection assembly;

[0041] 400a, irradiation module; 410a, lampshade; 411a, side cover; 412a, top cover; 420a, light source; 430a, support column;

[0042] 500a, imaging module;

[0043] 600a, data processing module;

[0044] B, microfluidic chip; 10b, chip body; 20b, injection chamber; 31b, quantitative chamber; 32b, amplification chamber; 33b, detection chamber; 34b, connection chamber; 41b, first waste liquid chamber; 42b, constant pressure flow channel; 50b, first siphon flow channel; 60b, distribution chamber; 70b, second waste liquid chamber; 80b, second siphon flow channel. DETAILED DESCRIPTION

[0045] The following embodiments of the present application are only used to illustrate the specific embodiments of the present application, and these embodiments cannot be understood as limiting the present application. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present application are deemed to be equivalent replacement methods and fall within the scope of protection of the present application.

[0046] Those skilled in the art should understand that, in the disclosure of the present application, the terms "first", "second", "third", "fourth", "fifth", etc. are only used to distinguish different structures, but do not limit the number, connection relationship, etc. of specific structures; in addition, the directions or positional relationships indicated by "longitudinal", "lateral", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. are based on the directions or positional relationships shown in the accompanying drawings, which are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation. Therefore, the above terms cannot be understood as limitations on the present application.

[0047] In this application, "proximal end" refers to the end of the centrifugal microfluidic chip that is close to the rotating shaft when it is set on the centrifugal nucleic acid detection device. "Distal end" refers to the end of the centrifugal microfluidic chip that is far away from the rotating shaft when it is set on the centrifugal nucleic acid detection device. During centrifugal detection, the liquid in the centrifugal microfluidic chip gradually moves from the proximal chamber to the distal chamber.

[0048] In one embodiment, Figures 1 to 4 As shown, a centrifugal nucleic acid detection device based on image analysis (hereinafter referred to as "device") is provided, comprising:

[0049] A frame, and the following structures arranged on the frame:

[0050] A rotating module, wherein the rotating module can carry the microfluidic chip and drive the microfluidic chip to rotate;

[0051] A temperature control module, which can perform temperature control (such as heating and / or maintaining a constant temperature) on the microfluidic chip carried by the rotating module so that the sample inside the microfluidic chip can undergo an amplification reaction (isothermal amplification RPA in this embodiment) and gene shearing (CRISPR shearing in this embodiment);

[0052] An irradiation module 400a, wherein the irradiation module 400a can emit light to stimulate the amplification reaction inside the microfluidic chip and the sample after gene shearing to emit fluorescence;

[0053] An imaging module 500a, when the sample after amplification reaction and gene shearing inside the microfluidic chip emits fluorescence, the imaging module 500a can shoot the area where the microfluidic chip B is set on the rotating module to obtain image data;

[0054] The data processing module 600a is electrically connected to the imaging module 500a to receive the image data and process the image data to obtain the nucleic acid detection result.

[0055] Of course, the data processing module 600a can also control the imaging module 500a to take pictures or stop taking pictures by being electrically connected to the imaging module 500a, so as to improve the automation level of the application equipment.

[0056] Regarding the rack, in order to facilitate the setting and layout of the various structures of the device of the present application, the rack of the present application includes a base plate 110a and a vertical plate 120a, wherein the imaging module 500a and the data processing module 600a are mainly arranged on the vertical plate 120a, and other structures are mainly arranged on the base plate 110a.

[0057] The material of the frame may be selected from existing metal materials, organic polymer materials, and inorganic materials. In the present application, metal materials and / or organic polymer materials are preferably included.

[0058] Regarding the microfluidic chip used in the device of the present application, those skilled in the art know that it should be a centrifugal microfluidic chip for nucleic acid detection, which can be an existing microfluidic chip or a microfluidic chip B developed by the applicant described below (structure reference Figure 5 ).

[0059] Regarding the number of microfluidic chips that can be carried on the rotating module, there can be more than one, preferably more than one (ie more than two, such as 2, 3, 4, 5, 6, 7, 8, 9 or more than 10).

[0060] Regarding the processing method of the image data by the data processing module 600a, the present application has no specific restrictions, as long as it can process the image data to obtain the nucleic acid detection results. Specifically, for example, a target detection algorithm (an algorithm that converts the detection problem into an image classification problem) is used to process the image data. In this embodiment, the Yolo algorithm in the target detection algorithm is used. Those skilled in the art know that before using a target detection algorithm (such as the Yolo algorithm) to process the image data, it is necessary to use a corresponding training set (such as a plurality of imaging modules 500a The image data obtained by shooting and the detection results of the samples corresponding to the image data by other means) and the like are trained for subsequent detection.

[0061] When using the above-mentioned device of the embodiment, first, the microfluidic chip is carried on the rotating module. The rotating module drives the microfluidic chip to rotate or stop rotating so that the sample flows in different cavities in the microfluidic chip B and mixes with the reagents in the cavity of the microfluidic chip. The temperature control module is cooperated to make the sample undergo amplification reaction and gene shearing (regarding the above-mentioned gene shearing (such as CRISPR cutting), technicians in this field can select appropriate reagents according to the sample and pre-set them in the detection cavity). After that, the irradiation module 400a emits light and irradiates the reacted sample to generate fluorescence, the imaging module 500a shoots the area where the microfluidic chip is set on the rotating module to obtain image data, and the data processing module 600a processes the image data to obtain the nucleic acid detection result.

[0062] As introduced in the background technology, in the prior art, in order to convert fluorescence or visible light into electrical signals through photoelectric sensors to display on the instrument reporting end, this solution has high requirements on the performance of photoelectric sensors, especially in the detection of multiple samples and multiple targets (such as multiple detection chambers involved in a microfluidic chip for reaction and / or multiple microfluidic chips), if the photoelectric sensors set by the instrument are less than the number of samples to be detected, the photoelectric sensors are required to detect the samples one by one, thereby reducing the detection efficiency; when the number of photoelectric sensors set corresponds to the number of samples, more photoelectric sensors need to be set, which undoubtedly increases the cost of the instrument, making it difficult to apply to instant detection scenarios with low cost requirements.

[0063] The above-mentioned device of this embodiment is a completely different technical solution from the above-mentioned prior art solution. It does not require a photoelectric sensor, but only requires a more mature, existing and lower-cost imaging module 500a to shoot the area where the microfluidic chip is set on the rotating module to obtain image data, and the data processing module 600a processes the image data to obtain the nucleic acid detection result. In particular, when multiple detection cavities are set on the microfluidic chip and / or multiple microfluidic chips are set on the device at the same time, the imaging module 500a can shoot the entire area where all microfluidic chips are set on the rotating module once to obtain image data, and the data processing module processes the image data once to obtain the detection results of all detection cavities in all microfluidic chips. Therefore, compared with the prior art, the above-mentioned device of this application has lower cost and greatly improves the detection efficiency.

[0064] In one embodiment, Figure 1 to Figure 3 As shown,

[0065] The rotation module comprises:

[0066] A motor 210a (a DC motor in this embodiment), wherein the motor 210a is directly or indirectly fixed to the frame;

[0067] The tray 220a is connected to the rotating shaft of the motor 210a so that the rotating shaft of the motor 210a can drive the tray 220a to rotate, and the tray can carry and fix more than one (preferably more than two, such as 2, 3, 4, 5, 6, 7, 8, 9 or more than 10) microfluidic chips.

[0068] Regarding the method of fixing the motor 210a to the frame, Figure 3 As shown, in this embodiment, the rotation module further includes a motor fixing seat 260a, and the motor fixing seat 260a is fixedly disposed with the bottom plate 110a, and the motor 210a is fixed to the motor fixing seat 260a with the rotating shaft facing upward.

[0069] Furthermore, the rotating module further includes: a tray fixing seat 230a, the tray fixing seat 230a is fixedly connected to the rotating shaft of the motor 210a, and the tray fixing seat 230a can be fixedly connected to the tray 220a.

[0070] This embodiment provides a structure of a rotating module, wherein the tray 220a carries and fixes the microfluidic chip, and the motor 210a drives the tray 220a to rotate, thereby achieving the purpose of the rotating module carrying the microfluidic chip and driving the microfluidic chip to rotate.

[0071] Furthermore, the rotation module further includes: a chip pressure cover 240a, the chip pressure cover 240a can press the microfluidic chip onto the tray 220a, so as to fix the microfluidic chip. Specifically, in this embodiment, the chip pressure cover 240a is provided with a through hole / blind hole corresponding to the rotating shaft of the motor, so that the chip pressure cover 240a is sleeved on the rotating shaft of the motor, and the chip pressure cover 240a presses the microfluidic chip onto the tray 220a by gravity. In addition, mutually matching protrusions and grooves are also provided between the tray 220a and the microfluidic chip and / or the chip pressure cover 240a, so that the motor can drive the microfluidic chip and / or the chip pressure cover 240a to rotate.

[0072] In one embodiment, Figure 3 As shown, the temperature control module includes:

[0073] A temperature-controlled tray assembly 310a, wherein the temperature-controlled tray assembly 310a is located on one side of the tray 220a;

[0074] The temperature control component 320a is disposed on a side of the temperature control tray component 310a facing the tray 220a, and the temperature control component 320a can control the temperature of the microfluidic chip (such as heating and / or maintaining a constant temperature).

[0075] The present application has no specific restrictions on the way in which the temperature control component performs temperature control, as long as it can achieve temperature control. In this embodiment, the temperature control component is a PI heating film.

[0076] That is, this embodiment provides a structure of a temperature control module, which carries a temperature control component 320a through a temperature control tray component 310a, and performs temperature control through the temperature control component 320a.

[0077] In addition, it should be noted that Figure 3 In the structure shown, the temperature control tray assembly 310a and the shaft of the motor 210a are not fixedly arranged. A through hole is arranged in the middle of the temperature control tray assembly 310a, and the shaft of the motor 210a passes through the through hole. The temperature control tray assembly 310a does not rotate with the rotation of the shaft of the motor 210a. When the shaft of the motor 210a and the tray 220a connected thereto stop rotating, the temperature control assembly 320a starts to perform temperature control.

[0078] Furthermore, the temperature control module further includes: a motion component 330a, which can drive the temperature control tray component 310a to approach or move away from the tray 220a. Thus, when the motor 210a shaft and the tray 220a connected thereto stop rotating, the motion component 330a drives the temperature control tray component 310a to approach the tray 220a to perform temperature control, thereby facilitating amplification reaction and gene shearing.

[0079] Specifically, regarding the motion component 330a, as Figure 1 As shown, it includes:

[0080] The motion unit 332a is directly or indirectly fixedly connected to the frame (such as Figure 3 As shown, in this embodiment, the motion component 330a further includes a fixed bracket 331a, the fixed bracket 331a is fixedly connected to the frame, and the motion unit 332a is fixedly connected to the fixed bracket 331a, that is, the motion unit 332a is indirectly fixedly connected to the frame);

[0081] The motion bracket 333a, the motion unit 332a can drive the temperature-controlled tray assembly 310a to move closer to or away from the tray through the motion bracket 333a.

[0082] Regarding the motion unit 332a, specific examples include a cylinder (at this time, those skilled in the art know that its fixed part is its housing, and its moving part is its push rod), an electric push rod, a lead screw, etc.

[0083] In this embodiment, Figure 1 As shown, the motion unit 332a used is a lead screw, wherein the housing of the drive motor of the lead screw is fixed to the fixed bracket 331a, and the motion bracket 333a is provided with an internal thread that cooperates with the screw of the lead screw, so that when the drive motor of the lead screw drives its screw to rotate, the motion bracket 333a is driven to move up and down, and the motion bracket 333a extends to the bottom of the temperature control tray assembly 310a or is fixed to the temperature control tray assembly 310a to drive the temperature control tray assembly 310a to move up and down. In addition, in order to prevent the motion bracket 333a from rotating, the fixed bracket 331a is further provided with a guide column parallel to the screw of the lead screw, and the motion bracket 333a is provided with a guide hole sleeved on the guide column.

[0084] In one embodiment, Figure 3 As shown, the temperature control tray assembly 310a includes:

[0085] First temperature controlled tray unit 311a;

[0086] A second temperature-controlled tray unit 312a, wherein the second temperature-controlled tray unit 312a is arranged in parallel with the first temperature-controlled tray unit 311a, and the second temperature-controlled tray unit 311a and the tray 220a are located on different sides of the first temperature-controlled tray unit 311a (in this embodiment, the tray 220a is located on the upper side of the first temperature-controlled tray unit 311a, and the second temperature-controlled tray unit 311a is located on the lower side of the first temperature-controlled tray unit 311a);

[0087] An elastic member 313a, wherein the elastic member 313a is disposed between the first temperature-controlled tray unit 311a and the second temperature-controlled tray unit 312a;

[0088] The motion component 330a can drive the second temperature-controlled tray unit 312a to move closer to or away from the tray 220a.

[0089] There is no specific limitation on the elastic member 313a in the present application, such as a compression spring or a rubber pad, etc. In the present embodiment, a compression spring is used.

[0090] When the motion component 330a drives the second temperature control tray unit 312a to approach the tray 220a, the second temperature control tray unit 312a presses the first temperature control tray unit 311a tightly against the lower part of the tray 220a through the elastic member 313a, so that the temperature control component 320 can be well close to the tray 220a for temperature control.

[0091] In one embodiment, Figure 1 , Figure 2 As shown, the temperature control module also includes:

[0092] The temperature detection component 340a can detect the temperature of the microfluidic chip.

[0093] In this embodiment, the temperature detection component 340a adopts non-contact temperature measurement, such as an infrared temperature sensor, so as to measure the temperature of the microfluidic line area on the upper surface of the tray 220a to improve the detection efficiency and facilitate the temperature control of the temperature control component.

[0094] In one embodiment, Figure 1 to Figure 3 As shown, the irradiation module 400a includes:

[0095] A lampshade 410a, wherein the lampshade 410a is disposed on the outer side of the tray 220a;

[0096] A plurality of light sources 420a are disposed inside the lampshade, and the light emitted by the light sources illuminates the microfluidic chip carried on the rotating module from the side.

[0097] like Figure 3 As shown, in the present application, the lampshade 410a includes a side cover 411a arranged around the outside of the tray 220a and a top cover 412a arranged on the upper part of the tray 220a. Of course, the middle of the top cover 412a is hollow to facilitate the temperature detection component 340a to detect the temperature. The light source 420a is arranged inside the side cover 411a.

[0098] Regarding the light source 420a, those skilled in the art can select a light source that emits light of a suitable wavelength to make it emit fluorescence according to the specific substance to be detected, which will not be described in detail here. In this embodiment, the light source 420a is a plurality of LED lamp beads, which are arranged in a ring inside the side cover 411a of the tray 220a.

[0099] In one embodiment, the imaging module includes: an image sensor (a CMOS image sensor in this embodiment); a filter that can filter out the light emitted by the illumination module and retain the fluorescence emitted by the amplification reaction inside the microfluidic chip and the sample after gene shearing; and a lens assembly.

[0100] Regarding filters, those skilled in the art can select appropriate filters according to the wavelength of the fluorescence excited by the specific substance to be detected and the wavelength of the light emitted by the light source, so as to filter out the light emitted by the light source and retain the fluorescence excited by the substance to be detected, which will not be elaborated here.

[0101] The image sensor acquires the filtered light (mainly fluorescence) and converts the light signal into a digital signal to obtain image data.

[0102] In one embodiment, a tray mark is provided on the tray (not shown in the figure, in this embodiment, it is an equilateral triangle mark corresponding to the setting position of the microfluidic chip);

[0103] The data processing module is at least capable of acquiring the position information of the microfluidic chip according to the information of the tray mark in the image data.

[0104] Thus, the data processing module can accurately identify the position of the microfluidic chip according to the tray mark, so as to further accurately identify the position of the detection cavity of the microfluidic chip, thereby facilitating data processing to obtain more accurate nucleic acid detection results.

[0105] In addition, the microfluidic chip is provided with a chip mark (not shown in the figure, in this embodiment, it is a character (such as numbers and / or letters) provided on different microfluidic chips, specifically a label with uppercase English letters (such as A / B / C / D, etc.));

[0106] The data processing module is at least capable of distinguishing different microfluidic chips according to the chip label information in the image data.

[0107] Thus, the data processing module can accurately identify different microfluidic chips based on the tray mark (such as the equilateral triangle mentioned above) and the chip mark (such as the uppercase English letter label attached to the microfluidic chip mentioned above), and further determine the positive and negative corresponding to each detection cavity on the chip based on the brightness of the chip detection cavity, so as to obtain the nucleic acid test results corresponding to different samples more accurately and efficiently.

[0108] In one embodiment, Figure 4 As shown, the data processing module 600a is also electrically connected to the rotation module to control the rotation module to drive the microfluidic chip to rotate. Specifically, the data processing module 600a can control the motor 210a to rotate or stop rotating.

[0109] In addition, the data processing module is also electrically connected to the temperature control module to control the temperature control module to perform temperature control. Specifically, the data processing module is electrically connected to the temperature detection component 340a to obtain the temperature detection result; the data processing module is electrically connected to the temperature control component 320a to perform temperature control, especially when the temperature control can be performed according to the detection result of the temperature detection component 340a, which can ensure more accurate temperature control; the data processing module is electrically connected to the motion component 330a (such as electrically connected to the drive motor of the lead screw) to control the temperature control component to be close to the tray 220a to facilitate heating, or to control the temperature control component to be away from the tray 220a to facilitate centrifugation.

[0110] The data processing module 600a can also be electrically connected to the irradiation module 400a to control the irradiation module to emit light to stimulate the amplification reaction and the sample after gene shearing to emit fluorescence, or control the light source to stop emitting light.

[0111] Those skilled in the art know that, based on the above-mentioned device provided in this application, the device and the corresponding microfluidic chip form a centrifugal nucleic acid detection system. That is, this application can also provide a centrifugal nucleic acid detection system, which includes: the above-mentioned centrifugal nucleic acid detection device based on image analysis; and, a microfluidic chip.

[0112] Regarding the microfluidic chip in the above centrifugal nucleic acid detection system, Figure 5 As shown, the applicant has designed the following microfluidic chip B after research, which includes:

[0113] A chip body 10b; and the following structure arranged on the chip body 10b:

[0114] Liquid injection chamber 20b;

[0115] Two or more detection modules, wherein the detection modules are arranged at the distal end of the injection chamber 20b, and the detection modules include a quantitative chamber 31b, an amplification chamber 32b, and a detection chamber 33b which are arranged and connected in sequence from the proximal end to the distal end, and the proximal end of the quantitative chamber 31b is connected to the distal end of the injection chamber 20b; and,

[0116] Constant pressure module;

[0117] Wherein, the constant voltage module comprises:

[0118] More than two first waste liquid chambers 41b, wherein the more than two first waste liquid chambers 41b are located at the far end of the chip body 10b;

[0119] A constant pressure channel 42b, wherein the constant pressure channel 42b is connected to each of the first waste liquid chambers 41b;

[0120] The proximal end of the injection cavity 20b and the proximal end of the cavity on the proximal side of the detection cavity 33b are both connected to the constant pressure module.

[0121] Regarding the chip body 10b, it may include a substrate and a cover layer covering the substrate, wherein each cavity and each flow channel are formed on the substrate, and the cover layer covers the substrate to seal each cavity and each flow channel on the substrate. Specifically, the liquid to be detected (i.e., the above-mentioned sample) can be added to the injection cavity 20b, the air-dried reagent / freeze-dried reagent for amplification can be added to the amplification cavity 32b, the probe reagent can be added to the detection cavity 33b, and then the substrate is covered with a cover layer for sealing, and then detected by the machine (the above-mentioned device).

[0122] Regarding the cavity on the proximal side of the detection cavity, those skilled in the art know that when the amplification cavity 32b is directly connected to the detection cavity 33b through a flow channel, the cavity on the proximal side of the detection cavity is the amplification cavity 32b; when, as described below, a connecting cavity 34b is further provided between the amplification cavity 32b and the detection cavity 33b, and the connecting cavity 34b is respectively connected to the amplification cavity 32b and the detection cavity 33b through the flow channel, that is, when the amplification cavity 32b is connected to the detection cavity 33b through the connecting cavity 34b, the cavity on the proximal side of the detection cavity is the connecting cavity 34b.

[0123] The present application has no specific restrictions on the shape of the microfluidic chip, but in order to facilitate the simultaneous arrangement of multiple groups of microfluidic chips in the above-mentioned device for centrifugal detection, the microfluidic chip is preferably fan-shaped. When the microfluidic chip is fan-shaped, multiple microfluidic chips can be arranged around the motor shaft of the above-mentioned device, so that the above-mentioned device performs the same treatment on multiple microfluidic chips, so as to perform the same detection on multiple microfluidic chips at the same time.

[0124] This embodiment provides a new microfluidic chip. When the microfluidic chip after adding the liquid to be detected and the detection reagent and sealing is set on the above device, the detection can be carried out through the following steps:

[0125] Step 1: Use centrifugal force to transfer the liquid to be tested from the injection chamber 20b to the quantitative chamber 31b. The quantitative chamber 31b can quantitatively separate the liquid in the injection chamber 20b. The volume of the quantitative chamber 31b determines the upper limit of the subsequent liquid transfer volume.

[0126] Step 2: Using centrifugal force to transfer the liquid to be detected from the quantitative chamber 31b to the amplification chamber 32b, then stop the centrifugation to perform amplification;

[0127] Step 3: After a preset time, restart the centrifugation to transfer the liquid in the amplification chamber 32b to the detection chamber 33b, and use an instrument to detect the fluorescence or visible light in the detection chamber 33b to obtain the nucleic acid detection result.

[0128] Among them, in the microfluidic chip provided by the present application, the first waste liquid chamber 41b and the constant pressure flow channel 42b in the constant pressure module can not only be used to collect waste liquid, but also, since in the process of liquid transfer from the proximal chamber to the distal chamber, the liquid will push the gas on its distal side to move toward the distal end. At this time, since the proximal end of the injection chamber 20b and the proximal end of the cavity on the proximal side of the detection chamber 33b are connected to the constant pressure module, the gas at the distal end of the liquid can flow back to the proximal side of the liquid through the first waste liquid chamber 41b and / or the constant pressure flow channel 42b of the constant pressure module, thereby ensuring that the liquid flows smoothly from the proximal end to the distal end when the microfluidic chip is completely sealed, and preventing the liquid to be tested from being contaminated due to the setting of vents during the detection process, thereby affecting the detection results.

[0129] In one embodiment, Figure 5 As shown, at least one opening is provided at the proximal end of the first waste liquid chamber 41b; more than two constant pressure flow channels 42b are connected to each of the first waste liquid chambers 41b through the openings of the first waste liquid chamber 41b to form a closed and surrounding constant pressure module; the injection chamber 20b and the detection module are arranged on the inner side of the constant pressure module.

[0130] Specifically, in this embodiment, when the microfluidic chip is fan-shaped, Figure 5 As shown, the first waste liquid chamber 41b and the constant pressure channel 42b are in a closed and surrounding shape arranged on the fan-shaped edge of the microfluidic chip, and other chambers are arranged therein, so that when multiple detection modules are arranged, the chambers of the detection modules can be conveniently connected with the first waste liquid chamber 41b and / or the constant pressure channel 42b of the constant pressure module, thereby facilitating the collection of waste liquid and balancing the internal pressure of the microfluidic chip.

[0131] In one embodiment, Figure 5 As shown, the distal end of the quantitative chamber 31b is connected to the proximal end of the amplification chamber 32b through the first siphon channel 50b; wherein the proximal end of the first siphon channel 50 is located on the proximal side of the proximal end of the quantitative chamber 31b.

[0132] This embodiment specifically provides a communication method between the distal end of the quantitative chamber 31b and the proximal end of the amplification chamber 32b, that is, communication through the first siphon channel 50b. When the proximal end of the first siphon channel 50b is located on the proximal side of the proximal end of the quantitative chamber 31b, during the centrifugation process of the above step 1, it can be ensured that the liquid in the quantitative chamber 31b will not easily flow to the chamber at its distal end. After stopping the centrifugation, in the above step 2, the first siphon channel 50b is used to attract the liquid to cross the proximal side of the first siphon channel 50b, and then the centrifugation is started, so as to transfer the liquid to be detected from the quantitative chamber 31b to the amplification chamber 32b.

[0133] In one embodiment, Figure 5 As shown, the microfluidic chip further includes the following structure arranged on the chip body 10b:

[0134] A distribution chamber 60b, wherein the distribution chamber 60b is located between the injection chamber 20b and the detection module;

[0135] A second waste liquid chamber 70b, wherein the second waste liquid chamber 70b is located at the distal end of the distribution chamber 60b;

[0136] Among them, the entrance of the distribution chamber 60b is connected with the injection chamber 20b; the quantitative chamber 31b and the second waste liquid chamber 70b in each of the detection modules are connected with the distribution chamber 60b, and the entrances of the second waste liquid chamber 70b and the distribution chamber 60b are located on different sides of the quantitative chamber of each of the detection modules so that the liquid flowing into the distribution chamber from the injection chamber first flows into the quantitative chamber in each of the detection modules and then flows into the second waste liquid chamber.

[0137] Then, in the above step 1, when the liquid is moved from the injection chamber 20b to its distal chamber by centrifugation, the liquid first enters the distribution chamber 60b through the inlet of the distribution chamber 60b, and then flows into the quantitative chamber 31b in each of the detection modules. After the quantitative chamber 31b is filled with liquid, the excess liquid will flow into the second waste liquid chamber 70b, thereby achieving the quantitative determination of the liquid to be detected and collecting the excess liquid (waste liquid).

[0138] In one embodiment, Figure 5As shown, the microfluidic chip further includes: a connecting chamber 34b, which is arranged between the amplification chamber 32b and the detection chamber 33b, and the connecting chamber 34b is connected to the amplification chamber 32b and the detection chamber 33b through a flow channel.

[0139] This embodiment provides a microfluidic chip in which a connecting cavity 34b is provided between the amplification cavity 32b and the detection cavity 33b.

[0140] In one embodiment, Figure 5 As shown, the distal end of the amplification chamber 32b is connected to the proximal end of the connection chamber 34b through a second siphon channel 80b; wherein the proximal end of the second siphon channel 80b is located on the proximal side of the proximal end of the amplification chamber 32b.

[0141] This embodiment specifically provides a way of connecting the distal end of the amplification chamber 32b and the proximal end of the connecting chamber 34b, that is, connecting through the second siphon channel 80b. In the above step 2, when the proximal end of the second siphon channel 80b is located on the proximal side of the proximal end of the amplification chamber 32b, during the centrifugation process, it can be ensured that the liquid in the amplification chamber 32b will not easily flow to the chamber at its distal end (connecting chamber 34b). After stopping the centrifugation, the second siphon channel 80b is used to attract the liquid to cross the proximal side of the second siphon channel 80b, and then the centrifugation is started, so as to transfer the liquid from the amplification chamber 32b to the connecting chamber 34b.

[0142] In one embodiment, Figure 5 As shown, the distal end of the connecting cavity 34b is connected to the detection cavity 33b through more than two flow channels.

[0143] When the distal end of the connecting cavity 34b is connected to the detection cavity 33b through only one flow channel, the gas in the detection cavity 33b makes it difficult for the liquid to enter the detection cavity 33b from the connecting cavity 34b. When the distal end of the connecting cavity 34b is connected to the detection cavity 33b through two or more flow channels, the liquid pressures of the two or more flow channels are different during the centrifugation process, so that the liquid can easily flow into the detection cavity 33b from part of the flow channels (such as one flow channel), and the gas in the detection cavity 33b can flow out of the detection cavity 33b from other flow channels (such as another flow channel), so as to facilitate the transfer of the liquid from the connecting cavity to the detection cavity 33b.

[0144] In one embodiment, Figure 5 As shown, the proximal end of the distribution chamber, the proximal end of the amplification chamber and / or the proximal end of the connection chamber are connected to the constant pressure module through a flow channel.

[0145] Through the configuration of this embodiment, it is possible to more conveniently ensure the pressure balance in the microfluidic chip, thereby facilitating the transfer of liquid from the proximal chamber to the distal chamber.

[0146] Although the embodiments of the present application are described above, the present application is not limited to the above specific embodiments and application fields, and the above specific embodiments are merely illustrative and instructive, rather than restrictive. A person of ordinary skill in the art can make many forms under the guidance of this specification and without departing from the scope of protection of the claims of the present application, all of which belong to the scope of protection claimed in the present application.

Claims

1. A centrifugal nucleic acid detection device based on image analysis, characterized in that: include: A frame, and the following structures arranged on the frame: A rotating module, wherein the rotating module can carry the microfluidic chip and drive the microfluidic chip to rotate; A temperature control module, which can control the temperature of the microfluidic chip carried by the rotating module so that the sample inside the microfluidic chip can undergo amplification reaction and gene shearing; An irradiation module, which can emit light to stimulate the samples undergoing amplification reaction and gene shearing inside the microfluidic chip to emit fluorescence; an imaging module, which can photograph the area on the rotating module where the microfluidic chip is arranged to obtain image data when the sample after amplification reaction and gene shearing inside the microfluidic chip emits fluorescence; and, A data processing module is electrically connected to the imaging module to receive the image data and process the image data to obtain a nucleic acid detection result.

2. The centrifugal nucleic acid detection device based on image analysis according to claim 1, characterized in that: The rotation module comprises: A motor, the motor being directly or indirectly fixed to the frame; The tray is connected to the rotating shaft of the motor so that the rotating shaft of the motor can drive the tray to rotate, and the tray can carry and fix more than one microfluidic chip.

3. The centrifugal nucleic acid detection device based on image analysis according to claim 2, characterized in that: The rotation module also includes: A tray fixing seat, wherein the tray fixing seat is fixedly connected to the rotating shaft of the motor, and the tray fixing seat can be fixedly connected to the tray.

4. The centrifugal nucleic acid detection device based on image analysis according to claim 2, characterized in that: The rotation module also includes: A chip pressing cover is provided, wherein the chip pressing cover can press the microfluidic chip onto the tray.

5. The centrifugal nucleic acid detection device based on image analysis according to claim 1, characterized in that: The temperature control module comprises: A temperature-controlled tray assembly, the temperature-controlled tray assembly being located on one side of the tray; A temperature control component is arranged on a side of the temperature control tray component facing the tray, and the temperature control component can control the temperature of the microfluidic chip.

6. The centrifugal nucleic acid detection device based on image analysis according to claim 5, characterized in that: The temperature control component is a PI heating film.

7. The centrifugal nucleic acid detection device based on image analysis according to claim 5, characterized in that: The temperature control module also includes: A motion component, wherein the motion component can drive the temperature-controlled tray component to move closer to or away from the tray.

8. The centrifugal nucleic acid detection device based on image analysis according to claim 7, characterized in that: The motion components include: A motion unit, the motion unit being directly or indirectly fixedly connected to the frame; The motion bracket is used for driving the temperature-controlled tray assembly to move closer to or away from the tray.

9. The centrifugal nucleic acid detection device based on image analysis according to claim 7, characterized in that: The temperature control tray assembly comprises: a first temperature controlled tray unit; a second temperature-controlled tray unit, wherein the second temperature-controlled tray unit is arranged in parallel with the first temperature-controlled tray unit, and the second temperature-controlled tray unit and the tray are located on different sides of the first temperature-controlled tray unit; an elastic member, wherein the elastic member is disposed between the first temperature-controlled tray unit and the second temperature-controlled tray unit; The motion component can drive the second temperature-controlled tray unit to move closer to or away from the tray.

10. The centrifugal nucleic acid detection device based on image analysis according to claim 5, characterized in that: The temperature control module also includes: A temperature detection component, wherein the temperature detection component can detect the temperature of the microfluidic chip.

11. The centrifugal nucleic acid detection device based on image analysis according to claim 2, characterized in that: The irradiation module comprises: A lampshade, the lampshade being arranged on the outer side of the tray; A plurality of light sources are arranged inside the lampshade, and the light emitted by the light sources irradiates the microfluidic chip carried by the rotating module from the side.

12. The centrifugal nucleic acid detection device based on image analysis according to claim 1, characterized in that: The imaging module includes: Image sensor; lens assembly; and, A filter is provided, wherein the filter can filter out the light emitted by the irradiation module and retain the fluorescence emitted by the amplification reaction inside the microfluidic chip and the sample after gene shearing.

13. The centrifugal nucleic acid detection device based on image analysis according to claim 1, characterized in that: The data processing module obtains the detection result based on the YOLO algorithm.

14. The centrifugal nucleic acid detection device based on image analysis according to claim 2, characterized in that: The pallet is provided with a pallet mark; The data processing module is at least capable of acquiring the position information of the microfluidic chip according to the information of the tray mark in the image data.

15. The centrifugal nucleic acid detection device based on image analysis according to claim 1, characterized in that: The microfluidic chip is provided with a chip mark; The data processing module is at least capable of distinguishing different microfluidic chips according to the chip label information in the image data.

16. The centrifugal nucleic acid detection device based on image analysis according to claim 1, characterized in that: The data processing module is also electrically connected to the rotation module to control the rotation module to drive the microfluidic chip to rotate; The data processing module is also electrically connected to the temperature control module to control the temperature control module to perform temperature control; and / or, The data processing module is also electrically connected to the irradiation module to control the irradiation module to emit light to stimulate the amplification reaction and the sample after gene shearing to emit fluorescence.

17. A centrifugal nucleic acid detection system, characterized in that: include: The centrifugal nucleic acid detection device based on image analysis according to any one of claims 1 to 16; and, Microfluidic chip.

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