Multi-sample multi-index quantity flexible switching microfluidic chip and intelligent detection system

Abstract

The application relates to a multi-sample multi-index quantity flexible switching microfluidic chip and an intelligent detection system, which comprises a chip body, a sample inlet pipeline, a plurality of sample addition holes arranged on the sample inlet pipeline, at least one exhaust pipeline arranged on the chip body, each exhaust pipeline being connected with the corresponding sample inlet pipeline through a connecting pipeline and a microcavity, at least one exhaust hole being arranged on each exhaust pipeline, and one or more than one isolation cavity being arranged on the flow channel between adjacent sample addition holes to form a simple valve with expansion material to realize flow path blocking and isolation. The simple valve is arranged on the flow channel, the microfluidic chip pipeline and the cavity are grouped through the on-off of the simple valve, the flexible change and switching of the sample quantity and the detection index quantity are realized, and the universality of the microfluidic chip is improved. Moreover, the intelligent detection system can realize the full integration of the raw sample inlet and the nucleic acid protein analysis result outlet.

Description

TECHNICAL FIELD

[0001] The present application relates to a multi-sample multi-index quantity flexible switching microfluidic chip and an intelligent detection system, and relates to the fields of biomedicine, clinical medicine, food safety and health and epidemic prevention. BACKGROUND

[0002] Nucleic acid protein detection analysis has been widely used in the fields of biomedicine, clinical medicine, food safety and health and epidemic prevention. The existing nucleic acid protein detection analysis method needs to rely on a plurality of instruments to be used in series to complete, and the detection time is long, about 2-3 days to report the results, the manual operation is very complex, the use of technical level and proficiency is required, the workload is large, and it must rely on a special molecular biology laboratory, and cannot be popularized and applied in primary medical units, communities, supermarkets, families and the like.

[0003] The microfluidic chip is a relatively advanced nucleic acid protein detection analysis method in recent years, and the existing microfluidic chip structure is often relatively special and fixed. The number of samples and the number of detection indexes corresponding to the added samples are fixed with the microfluidic chip structure, and cannot be expanded, and cannot adapt to new detection changes. Moreover, the microfluidic chip is relatively precise, and needs to be precisely processed and molded to ensure precision, and the cost is relatively high. Once the microfluidic chip does not have universality, it will bring limitations to the use of the microfluidic chip kit, and the corresponding production cost is higher, which is not suitable for the popularization and application of the microfluidic chip technology. It is also the fundamental reason why the cost of nucleic acid protein detection based on the microfluidic chip is high at the present stage, and the detection cost cannot be greatly reduced. SUMMARY

[0004] The present application aims to at least solve one of the technical problems existing in the prior art. To this end, in view of the above problems, the purpose of the present application is to provide a multi-sample multi-index quantity flexible switching microfluidic chip and an intelligent detection system, which realizes flexible switching between the number of samples and the number of detection indexes on the microfluidic chip, greatly reduces manual operation, and effectively avoids cross contamination and infection.

[0005] In order to achieve the above-mentioned application purpose, the technical scheme adopted by the present application is:

[0006] In a first aspect, the present application provides a multi-sample multi-index quantity flexible switching microfluidic chip, which comprises:

[0007] A chip body;

[0008] A sample inlet pipeline, the sample inlet pipeline is arranged on the chip body, and a plurality of sample addition holes are arranged on the sample inlet pipeline;

[0009] An exhaust pipe is arranged on the chip body, each of the exhaust pipes is connected with the corresponding sample inlet pipe through a connecting pipe and a micro-cavity, wherein two ends of a micro-cavity are communicated with the sample inlet pipe and the exhaust pipe through a connecting pipe to form a detection channel, and the number of the detection channels is set to be several; at least one exhaust hole is arranged on each of the exhaust pipes.

[0010] An isolation cavity is arranged on the flow channel between adjacent sample wells, and each of the isolation cavities is provided with a simple valve to realize flow path blocking and isolation.

[0011] The microfluidic chip further comprises an expansion material or a microstructure, wherein the expansion material expands to form the simple valve to realize flow path blocking and isolation, or the microstructure deforms the isolation cavity pipe through pressurization, hot pressing or ultrasonic to form the simple valve to realize flow path blocking and isolation.

[0012] The microfluidic chip further comprises a straight line or a curved shape, and two ends of the sample inlet pipe are respectively provided with a sample well, and a plurality of sample wells are arranged at intervals in the middle of the sample inlet pipe according to needs.

[0013] The microfluidic chip further comprises a water-swelling material, and the water-swelling material comprises starch grafted acrylate polymer cross-linking material, acrylamide-acrylate copolymer cross-linking material and / or hydrogel resin, and when the sample inlet pipe is added with a sample, the expansion material swells to form a simple valve.

[0014] The microfluidic chip further comprises a heating expansion material, and the heating expansion material comprises liquid metal, memory alloy, heat-expanding high polymer material and / or heat-expanding polymer gel, and when the pipe is heated to a set temperature, the expansion material swells to form a simple valve.

[0015] The microfluidic chip further comprises one or more isolation cavities arranged on the flow channel, including the isolation cavities arranged on the sample inlet pipe and / or the connecting pipe, wherein the isolation cavities are arranged in the following forms:

[0016] The isolation cavities are arranged on the sample inlet pipe.

[0017] The isolation cavities are arranged on the connecting pipe, including that the isolation cavities are arranged at a tangent position of any connecting pipe, the isolation cavities are arranged at a position where the connecting pipe is partially intersected, or the isolation cavities are connected with the connecting pipe through a pipe section.

[0018] The microfluidic chip further comprises a waterproof and breathable film covering the exhaust hole, and gas is exhausted from the exhaust hole through the exhaust pipeline; and / or a sealing silicone rubber pad covering the sample loading hole to prevent sample contamination.

[0019] In a second aspect, the present application further provides an intelligent detection system comprising the microfluidic chip, the injection tube type sample preparation unit, the reagent storage unit, the optical detection system, the temperature control system, the heating unit, the heating and oscillation unit, the motion control system and / or the intelligent analysis system.

[0020] The injection tube type sample preparation unit is configured to perform lysis and extraction, purification and concentration of nucleic acid proteins on a sample.

[0021] The reagent storage unit is configured to provide reagent storage for sample preparation of the injection tube type sample preparation unit and reaction of the microfluidic chip.

[0022] The optical detection system is configured to detect the results of the reaction on the microfluidic chip.

[0023] The temperature control system is configured to perform constant or variable temperature heating and temperature control on the sample preparation process in the injection tube type sample preparation unit, and / or constant or cyclic variable temperature heating and temperature control on the reaction process of the microfluidic chip.

[0024] The heating and oscillation unit is configured to oscillate and / or heat the injection tube type sample preparation unit.

[0025] The heating unit is configured to heat and / or cool the microfluidic chip.

[0026] The motion control system is configured to control the sample preparation process in the injection tube type sample preparation unit, and / or to control the motion of the microfluidic chip during sample loading and / or reaction process.

[0027] The intelligent analysis system is configured to perform human-computer interaction control on the sample preparation process in the injection tube type sample preparation unit and / or the reaction process of the microfluidic chip, and / or to store, analyze, display and / or early warning report the sample reaction detection results obtained by the optical detection system.

[0028] The intelligent detection system further comprises an injection tube barrel, a piston, a filter membrane or filter paper, a sealing ring, a needle and a protective sleeve, wherein the piston is movably arranged in the injection tube barrel, the filter membrane or filter paper and the sealing ring are arranged at the bottom of the injection tube barrel, the needle is arranged at the bottom outlet of the injection tube barrel, and the protective sleeve is arranged on the needle.

[0029] The intelligent detection system, further, the injection tube cylinder pre-stores a swab storage elution lysis liquid and / or a magnetic bead, a glass bead, a nanomaterial or a polymer material after surface modification treatment, wherein the surface modification treatment comprises anion exchange broad-spectrum modification of negative electricity in an acidic environment, specific group modification for target capture based on nucleic acid hybridization or protein immune combination principle, adsorption of negative nucleic acid protein in an acidic environment, release of negative nucleic acid protein in an alkaline environment, adsorption of nucleic acid protein at low temperature and / or release of nucleic acid protein at high temperature.

[0030] The intelligent detection system, further, the microfluidic chip, the injection tube sample preparation unit, the reagent storage unit, the optical detection system, the temperature control system, the heating unit, the heating oscillation unit, the motion control system and the intelligent analysis system are all used in combination to realize full integration of raw sample input and nucleic acid protein analysis result output; or the injection tube sample preparation unit, the reagent storage unit, the temperature control system, the heating oscillation unit, the motion control system and the intelligent analysis system are used in combination to form an independent sample processing system for sample lysis and nucleic acid protein extraction, purification and concentration; or the microfluidic chip, the optical detection system, the temperature control system, the heating unit, the motion control system and the intelligent analysis system are used in combination to form an independent nucleic acid protein detection intelligent analysis system.

[0031] The present application has the following characteristics due to the above technical solutions:

[0032] 1. Compared with the conventional microfluidic chip method, the simple valve is arranged on the flow channel, the microfluidic chip pipeline and cavity are grouped by the on-off of the simple valve, the flexibility of sample quantity and detection index quantity is realized, the versatility of the microfluidic chip is improved, the technical use threshold and the requirement for site environmental conditions are reduced, and the popularization and application needs of microfluidic chip precision medicine molecular diagnosis are met.

[0033] 2. Compared with the conventional centrifugal tube or hole plate biochemical analysis method, the present application not only greatly reduces the use consumption of sample reagent (can be <1 muL / index), can detect dozens or even hundreds of specific gene protein analysis indexes in parallel once, greatly reduces the manual operation steps, reduces the labor intensity, makes the swab sampling detection more safe, robust and efficient; in addition, the user can automatically perform nucleic acid amplification or protein immune detection analysis, without the help or assistance of professional technicians, and without site environmental requirements, can realize standardized operation and use in the field, at home, in the community, in medical units at all levels, in health and epidemic prevention departments, in food safety departments and the like, and avoid the influence of human factors.

[0034] 3. Compared with conventional swab elution methods, this invention is compatible with two methods: sealed elution separation of sampled and adsorbed samples by adding the elution solution to the sampling swab or by directly adding the sampling swab. It efficiently transfers the elution solution in situ. After sampling, the sample is placed in an injection tube and the cap is not opened again, ensuring a completely sealed process. This effectively avoids serious problems such as infection of subsequent operators and cross-contamination throughout the process, which cannot guarantee safety and reliability.

[0035] In summary, this invention can effectively improve the versatility of microfluidic chips, solve the problem of advanced diagnostic technology and equipment needs in smart healthcare for convenient public services, and realize the forward shift of health and epidemic prevention and precision medicine from large hospitals and third-party testing centers to townships, communities, and homes. This effectively alleviates the pressure on large hospitals and enables intelligent networks for medical and health epidemic prevention to cover townships, communities, homes, large hospitals, third-party testing centers, and health and epidemic prevention departments. It effectively improves the advanced scientific and technological level of precision medicine and health and epidemic prevention for the entire population and can be widely applied in field, home, community, township clinics, physical examination centers, and clinical medical units at all levels, health and epidemic prevention departments, food safety management departments, as well as biomedical research, biochemical warfare agent testing, environmental monitoring, or other fields. Attached Figure Description

[0036] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Throughout the drawings, the same reference numerals denote the same parts. In the drawings:

[0037] Figure 1 This is a schematic diagram of the structure of a microfluidic chip that allows for flexible switching of the number of multiple samples and multiple indicators according to an embodiment of the present invention.

[0038] Figure 2 This is a schematic diagram of the isolation chamber structure of a simplified valve according to an embodiment of the present invention.

[0039] Figure 3 This is a schematic diagram of the microstructure of a simplified valve according to an embodiment of the present invention.

[0040] Figure 4 This is a schematic diagram of the structure of an intelligent detection system according to an embodiment of the present invention.

[0041] Figure 5 This is a schematic diagram of the structure of an injection tube sample preparation unit according to an embodiment of the present invention.

[0042] Figure 6 This is a schematic diagram of the structure for preparing an independent nucleic acid / protein extraction, purification, and concentration sample according to an embodiment of the present invention. Detailed Implementation

[0043] It is to be understood that the terminology used herein is for the purpose of describing particular example embodiments only and is not intended to be limiting. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. The terms "comprises", "comprising", "includes", "including" and "has" are inclusive and therefore specify the presence of stated features, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and / or groups thereof. The method steps, processes, and operations described herein are not to be construed as necessarily requiring their performance in the particular order

[0044] Although the terms first, second, third, etc. can be used herein to describe various elements, components, regions, layers and / or sections, these elements, components, regions, layers and / or sections should not be limited by these terms. These terms can be only used to distinguish one element, component, region, layer or section from another region, layer or section. Terms such as "first", "second", and other numerical terms when used herein do not imply a sequence or order unless clearly indicated by the context. Thus, a first element, component, region, layer or section discussed below could be termed a second element, component, region, layer or section without departing from the teachings of the example embodiments.

[0045] Spatially relative terms, such as "inner", "outer", "beneath", "below", "lower", "above", "upper", and the like, can be used herein for ease of description to describe one element or feature's relationship to another element(s) or feature(s) as illustrated in the figures. The spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientations depicted in the figures.

[0046] Because the existing microfluidic chip does not have universality, the production cost is relatively high, and the nucleic acid protein detection based on the microfluidic chip is high in cost. The present application provides a multi-sample multi-index quantity flexible switching microfluidic chip, which comprises a chip body; a sample inlet pipeline is arranged on the chip body, and a plurality of sample addition holes are arranged on the sample inlet pipeline; at least one exhaust pipeline is arranged on the chip body, each exhaust pipeline is connected with the corresponding sample inlet pipeline through a connecting pipeline and a microcavity, wherein the two ends of a microcavity are communicated with the sample inlet pipeline and the exhaust pipeline through a connecting pipeline to form a detection channel, and the number of detection channels is set to be several; at least one exhaust hole is arranged on each exhaust pipeline; one or more isolation cavities are arranged on the flow channel between adjacent sample addition holes, and an expansion material or a microstructure is arranged in each isolation cavity, wherein the expansion material forms a simple valve after expansion to realize flow path blocking and isolation, and the microstructure deforms the microstructure of the isolation cavity and the pipeline through pressurization, hot pressing or ultrasonic wave to form a simple valve to realize flow path blocking and isolation. Moreover, the intelligent detection system of the present application can realize the full integration of the original sample inlet and the nucleic acid protein analysis result outlet; it can also be simplified into a sample lysis and nucleic acid protein extraction and purification concentrated independent sample processing system; it can also be compatible with nucleic acid proteins extracted by other sample preparation methods to form an independent nucleic acid protein detection intelligent analysis system. Compared with the conventional nucleic acid amplification / protein immune analysis microfluidic chip method, the simple valve type flow path isolation technology of the present application can realize the flexible switching between the sample quantity and the detection index quantity, and the injection tube type full integration nucleic acid / protein extraction and purification method can realize the nucleic acid amplification / protein immune analysis of two systems sharing one set of hardware platform. When used, the user only needs to add the original sample, and the intelligent detection system can complete the full integration of the nucleic acid amplification / protein immune reaction detection analysis of the sample inlet and the result outlet, without complex professional technical operation and without special laboratory site environment requirement, which can meet the needs of on-site, home, community, medical center, medical units at all levels, health and epidemic prevention departments and food safety management departments, as well as nucleic acid / protein analysis standardized operation in the field of biomedical research, biochemical war agent detection, environmental detection or other fields.

[0047] Exemplary embodiments of the present application will be described in greater detail below with reference to the accompanying drawings. While exemplary embodiments of the present application are shown in the drawings, it is understood that the present application can be embodied in various forms and should not be limited by the embodiments set forth herein. Rather, these embodiments are provided so that this application will be thoroughly and completely understood, and will fully convey the scope of the application to those skilled in the art.

[0048] It should be noted that each structure of the embodiment can be supported and fixed by a corresponding structure, and the specific fixing mode is not limited, as long as it can meet the corresponding functions of the application. In order to facilitate understanding, convenient use, reduce weight and other requirements, the drawings of the embodiment mainly adopt hollow cylinders, rectangles, squares, circles or ellipses for module schematic implementation, which is only an example and is not limited thereto. Only one or more repeated units are drawn, which is only an example and is not limited thereto. The actual implementation can be repeated by using a corresponding structure.

[0049] Embodiment one: as shown in the embodiment, the multi-sample multi-index quantity flexible switching microfluidic chip provided by the embodiment comprises: Figure 1

[0050] The chip body;

[0051] The chip body is provided with a linear or curved sample inlet pipeline, and a plurality of sample addition holes are arranged at intervals on the sample inlet pipeline. The embodiment is described by taking an S-shaped sample inlet pipeline 1 as an example, which is only an example and is not limited thereto. The two ends of the S-shaped sample inlet pipeline 1 are respectively provided with a sample addition hole 11, 16, and the bending portions of the S-shaped sample inlet pipeline 1 are each provided with a sample addition hole 12-15. The embodiment is provided with six sample addition holes 11-16, which is only an example and is not limited thereto. The position and number of sample addition holes can be set according to actual needs.

[0052] The chip body is further provided with at least one exhaust pipeline 2, and the embodiment is provided with two exhaust pipelines 21, 22, which is only an example and is not limited thereto. The number of exhaust pipelines can be set according to actual detection needs. Each exhaust pipeline is connected to a corresponding S-shaped sample inlet pipeline 1 through a connecting pipeline 5 and a microcavity. One end of a microcavity is connected to a sample inlet pipeline and the other end is connected to an exhaust pipeline through a connecting pipeline 5 to form a detection channel. The number of detection channels can be set according to actual needs. In the embodiment, there are 40 detection channels, which is only an example and is not limited thereto. Each exhaust pipeline 21(22) is provided with at least one exhaust hole 23(24). Among them, the flow channel between adjacent sample addition holes, such as the sample inlet pipeline or the connecting pipeline, can be provided with one or more isolation cavities. Each isolation cavity is provided with an expansion material or a microstructure. The expansion material expands to form a simple valve for realizing flow path blocking and isolation. The microstructure deforms the isolation cavity microstructure and the pipeline to form a simple valve to realize flow path blocking and isolation by pressurization, hot pressing or ultrasonic waves. Therefore, the embodiment adopts a simple valve flow path isolation technology. The simple valve is arranged on the flow channel. The on-off of the simple valve groups the microfluidic chip pipeline and the microcavity, so as to realize the flexible change and switching of the number of samples and detection indexes.

[0053] ​In a preferred embodiment of the present application, the swelling material can be a water-swellable material, such as starch grafting acrylate polymer cross-linking material, acrylamide-acrylate copolymer cross-linking material, hydrogel resin, etc. When the sample is added to the sample inlet pipe, the swelling material swells in water to form a simple valve, thereby achieving flow path blocking and isolation.

[0054] Further, the isolation cavity can be arranged at any of the following positions on the flow channel:

[0055] As shown in Figure 2 (a), the isolation cavity can be arranged on the sample inlet pipe, for example, the sample inlet pipe between adjacent sample wells is provided with an isolation cavity 31-35. Each isolation cavity is provided with, for example, swelling material to form a simple valve, which can achieve rapid flow path blocking and isolation, and ensure that after different samples enter from different sample wells, they can only enter the corresponding microcavity region and cannot spread and contaminate other microcavity regions.

[0056] The isolation cavity can be arranged on the connecting pipe, as shown in Figure 2 (b), the isolation cavity can be arranged at any tangent position of the connecting pipe; as shown in Figure 2 (c), the isolation cavity can be arranged at a position intersecting with part of the connecting pipe. As shown in Figure 2 (d), the isolation cavity can be connected to the connecting pipe through a section of pipe. Therefore, the swelling material can be arranged in the isolation cavity beside the connecting pipe. By changing the distance or the thickness of the pipe between the connecting pipe and the isolation cavity, the swelling material can swell in water or by heating to form a simple valve, which can achieve several seconds to several minutes of delayed flow path blocking and isolation, and ensure that the added sample completely fills the microcavity and expels all gas.

[0057] In a preferred embodiment of the present application, the microstructure can be subjected to pressurization, heat pressing or ultrasonic treatment, etc. to deform the microstructure and the pipe to form a simple valve, thereby achieving flow path blocking and isolation. As shown in Figure 3 , the microstructure is not limited to any shape and can be arranged as needed, for example, spherical, triangular, zigzag, etc.

[0058] In a preferred embodiment of the present application, the swelling material can also be a heating swelling material placed in the isolation cavity, such as liquid metal, memory alloy, heat-expandable high polymer material, heat-expandable polymer gel, etc. When the pipe is heated to a certain temperature, the swelling material swells by heating to form a simple valve, thereby achieving flow path blocking and isolation.

[0059] Further, the exhaust hole 23(24) can be covered with a waterproof and breathable film, and the gas is discharged from the exhaust hole 23(24) through the exhaust pipe 21(22).

[0060] Further, the sample well 11-16 is covered with a sealing silicone rubber pad to prevent sample contamination.

[0061] In summary, when the multi-sample multi-index quantity flexible switching microfluidic chip of the embodiment is used, the simple valves in the embodiment are arranged in the isolation cavities 31-35 and 36-37, and the nucleic acid / protein analysis indexes are arranged in the microcavities 401-440 by low-melting-point agarose embedding or freeze-dried microsphere storage; the sample enters the sample inlet channel 1 from the sample inlet holes 11-16, fills the microcavities 401-440 quickly through the connecting channel 5, and is then blocked and isolated by the simple valves of the isolation cavities arranged beside the connecting channel 36-37 or on the sample inlet channel 31-35 to prevent cross contamination. For example, when all the simple valves of the isolation cavities 31-35 are placed in water-expanding or heat-expanding materials, the microcavity area is divided into six groups, corresponding to the six sample inlet holes 11-16, and six samples can be input; the sample input by the sample inlet hole 11 corresponds to the microcavities 401-405 for detecting five indexes in total, the sample input by the sample inlet hole 12 corresponds to the microcavities 406-410 for detecting five indexes in total, the sample input by the sample inlet hole 14 corresponds to the microcavities 411-415 and 421-425 for detecting ten indexes in total, the sample input by the sample inlet hole 13 corresponds to the microcavities 416-420 and 426-430 for detecting ten indexes in total, the sample input by the sample inlet hole 15 corresponds to the microcavities 431-435 for detecting five indexes in total, and the sample input by the sample inlet hole 16 corresponds to the microcavities 436-440 for detecting five indexes in total. For another example, when only some of the simple valves of the isolation cavities 31-35, such as 32 and 34, are placed in water-expanding or heat-expanding materials, the microcavity area is divided into three groups, and only three sample inlet holes 11 or 12, 13 or 14, 15 or 16 are needed to correspond to three samples; the sample input by the sample inlet hole 11 or 12 corresponds to the microcavities 401-410 for detecting ten indexes in total, the sample input by the sample inlet hole 13 or 14 corresponds to the microcavities 411-430 for detecting twenty indexes in total, and the sample input by the sample inlet hole 15 or 16 corresponds to the microcavities 431-440 for detecting ten indexes in total. Therefore, based on the arrangement of the simple valves of the isolation cavities 31-35, the flexible switching combination of one to six different sample quantities and five to forty different detection index quantities can be realized. Moreover, with the extension of the sample inlet channel and the increase of the number of microcavities, the changes of the sample quantity and the detection index quantity will be more abundant, and the needs of different actual applications for the changes of the sample quantity and the detection index quantity can be met.

[0062] Embodiment Two: The embodiment provides an injection tube type sample preparation fully integrated nucleic acid amplification or protein immune reaction intelligent detection system, which realizes the input of one or more mixed multiple samples containing but not limited to pathogenic bacteria, microorganisms, tissue cells, exosomes, and exovesicles in a biological sample and the output of the detection results of multiple nucleic acid / protein analysis indexes.

[0063] As Figure 4As shown, the injection tube type sample preparation full integration nucleic acid amplification or protein immune reaction intelligent detection system provided by the embodiment includes the multi-sample multi-index quantity flexible switching microfluidic chip 10 described in embodiment one, one or more nucleic acid / protein extraction and purification injection tube type sample preparation units 20, one or more reagent storage units 30, an optical detection system 40, a temperature control system 50, a heating unit 60, a heating and oscillation unit 70, a motion control system 80, and / or an intelligent analysis system 90.

[0064] The injection tube type sample preparation unit 20 is configured to perform automatic nucleic acid / protein extraction and purification and concentration processing on the original sample in one injection tube by combining one or more methods of high temperature and / or oscillation crushing and / or chemical reagent lysis;

[0065] The reagent storage unit 30 is configured to set multiple reagent and waste liquid storage cavities according to the nucleic acid / protein sample preparation and amplification / immune reaction needs, store corresponding reagents inside, and seal the cavities by rubber plugs, which are suitable for sampling or discharging waste liquid by inserting a syringe needle.

[0066] The optical detection system 40 is configured to detect the results of nucleic acid amplification or protein immune reaction on the multi-sample multi-index quantity flexible switching microfluidic chip 10.

[0067] The temperature control system 50 is configured to perform constant temperature or variable temperature heating temperature control on the sample preparation process in the injection tube type nucleic acid / protein extraction and purification sample preparation unit, and / or perform constant temperature or cyclic variable temperature heating temperature control on the nucleic acid amplification or protein immune reaction process of the multi-sample multi-index quantity flexible switching microfluidic chip 10.

[0068] The heating unit 60 is configured to heat and / or cool the multi-sample multi-index quantity flexible switching microfluidic chip 10.

[0069] The heating and oscillation unit 70 is configured to oscillate and / or heat the injection tube type sample preparation unit 20.

[0070] The motion control system 80 is configured to perform oscillation crushing control, liquid suction / discharge control, etc. on the sample preparation process in the injection tube type nucleic acid / protein extraction and purification sample preparation unit, and / or perform sample addition and nucleic acid amplification or protein immune reaction process motion control, etc. on the multi-sample multi-index quantity flexible switching microfluidic chip 10.

[0071] The intelligent analysis system 90 is configured to perform human-computer interaction control on the nucleic acid / protein extraction, purification, and concentration sample preparation process and the detection process of nucleic acid amplification or protein immune reaction of the biological sample, and / or store, analyze, display, and / or early warning and direct reporting of the detection results of nucleic acid amplification or protein immune reaction of the biological sample.

[0072] In one preferred embodiment of the present application, as shown in Figure 5 The injection tube type sample preparation unit of the present embodiment is provided as two parallel sample preparation units, and one of the injection tube type sample preparation units 20 is taken as a specific embodiment for illustration, which includes an injection tube 201, a piston 202, a filter membrane or filter paper 203, a sealing ring 204, a needle 205 and a protective sleeve 206. The piston 202 is movably arranged in the injection tube 201, the filter membrane or filter paper 203 and the sealing ring 204 are arranged at the bottom of the injection tube 201, the needle 205 is arranged at the bottom outlet of the injection tube 201, and the protective sleeve 206 is arranged on the needle 205.

[0073] Further, the injection tube 201 is preloaded with a sample inactivation and nucleic acid protein degradation prevention lysis preservation solution. The piston 202 is first removed, and a swab sampling or minimally invasive sampling method is used to add one or more mixed original samples, such as but not limited to pathogenic bacteria, microorganisms, tissue cells, exosomes, exovesicles, etc., such as swab wash, biopsy puncture liquid, saliva sputum, sweat, blood, urine, feces, etc., into the preservation solution in the injection tube 201. The piston 202 is then installed and adjusted to the appropriate position, and then installed into the injection tube type sample preparation full integration nucleic acid amplification or protein immune reaction intelligent detection system. Through the automatic control of the injection tube type sample preparation full integration nucleic acid amplification or protein immune reaction intelligent detection system, the "sample in, result out" full automation nucleic acid amplification or protein immune reaction precision medicine molecular diagnosis can be realized.

[0074] Further, the injection tube 201 is pre-stored with a swab preservation elution lysis solution and / or magnetic beads or glass beads or other nanomaterials or high polymer materials after surface modification treatment. The surface modification treatment includes but is not limited to anion exchange broad-spectrum modification under acidic environment, specific group modification based on nucleic acid hybridization or protein immune binding principle for target grabbing, etc. The nucleic acid protein or other biological samples are adsorbed under acidic environment, released under alkaline environment, or adsorbed under low temperature and released under high temperature. Therefore, the present application uses filter membrane or filter paper and / or magnetic beads or anion exchange adsorption nucleic acid protein concentration and purification strategies, which not only is suitable for nucleic acid protein detection and analysis of different biological samples such as pathogenic bacteria, microorganisms, tissue cells, exosomes, exovesicles, etc., but also can effectively reduce the amount of elution solution and increase the initial concentration of the recovered adsorbed sample or lysis nucleic acid protein by nearly 100 times for subsequent use.

[0075] In one preferred embodiment of the present application, as shown in Figure 6As shown, the injection tube type sample preparation unit 20, under the joint action of the temperature control system 50, the heating and shaking unit 70 and the motion control system 80, produces biological sample cracking in an independent or combined manner of mechanical crushing and high-temperature cracking, and can also produce biological sample cracking in one or a combination of high temperature, mechanical crushing and chemical reagent cracking in combination with the preloaded chemical cracking reagent in the injection tube, so that the original biological sample in the injection tube releases nucleic acid protein, and under the action of the motion control system 80, realizes sequential absorption of various reagents from the reagent storage unit 30 and discharge of waste liquid generated in the intermediate processing process to the waste liquid storage cavity of the reagent storage unit 30, realizes automatic nucleic acid / protein extraction, purification and concentration processing of the original sample in one injection tube, and finally injects the purified and concentrated nucleic acid / protein into the multi-sample multi-index quantity flexible switching microfluidic chip 10 or other nucleic acid protein detection system, to meet the sample preparation application needs of automatic nucleic acid / protein extraction, purification and concentration processing of the original sample.

[0076] In a preferred embodiment of the present application, the reagent storage unit 30 is provided with a plurality of reagent storage cavities and at least one large-volume waste liquid storage cavity, which is sealed with a rubber plug and is suitable for insertion of a syringe needle for sample absorption or waste liquid discharge, to meet the needs of nucleic acid / protein extraction, purification and concentration processing of biological samples.

[0077] In a preferred embodiment of the present application, the heating and shaking unit 70 is used for sample cracking processing of the original biological sample in the injection tube barrel 201 in one or a combination of high temperature and / or shaking crushing and / or chemical reagent cracking, to efficiently separate and produce nucleic acid protein.

[0078] In a preferred embodiment of the present application, the optical detection system 40 uses existing technologies to detect the results of nucleic acid amplification or protein immune reaction, including an objective lens, an illumination light source, an excitation filter, an imaging lens, an emission filter and a detector, which can detect fluorescent signals through filter switching, can measure white light interference hyperspectral signal thickness decoding super-resolution by placing the filter in a vacant position, and can use a mobile phone or human eye for turbidity, phase contrast, chroma, spectrum or other spectrophotometric measurements, which will not be described here.

[0079] In a preferred embodiment of the present application, the temperature control system 50 is combined with the heating unit 60 to realize constant temperature or cyclic temperature control of the nucleic acid amplification or protein immune reaction process of the multi-sample multi-index quantity flexible switching microfluidic chip 10, to meet the needs of nucleic acid constant temperature or cyclic temperature amplification or protein immune reaction.

[0080] Further, the temperature control system 50 cooperates with the heating and oscillation unit 70 to realize constant or variable temperature heating control of the sample preparation process in the injection tube type nucleic acid / protein extraction and purification sample preparation unit, meeting the sample preparation needs of nucleic acid / protein extraction and purification.

[0081] In a preferred embodiment of the present application, the motion control system 80 cooperates with the heating and oscillation unit 70 to realize oscillation and crushing control, liquid suction and discharge control, etc. of the sample preparation process in the injection tube type nucleic acid / protein extraction and purification sample preparation unit.

[0082] Further, the motion control system 80 cooperates with the optical detection system 40 to realize motion control of the nucleic acid amplification or protein immune reaction process in the multi-sample and multi-index quantity flexible switching microfluidic chip 10, meeting the needs of imaging or confocal scanning detection of the real-time signal of the nucleic acid amplification or protein immune reaction in the microcavity of the multi-sample and multi-index quantity flexible switching microfluidic chip 10.

[0083] In a preferred embodiment of the present application, the intelligent analysis system 90 includes a microprocessor signal storage processing and analysis unit, a wired or wireless communication interface, a display terminal, and a cloud big data server, etc., cooperates with the temperature control system 50, the motion control system 80 and the optical detection system 40 to realize human-computer interaction control of the biological sample preparation process and the nucleic acid amplification or protein immune reaction detection process, and stores, analyzes, displays and / or early warns and reports the detection results, wherein,

[0084] The signal storage processing and analysis unit is provided with a big data prior knowledge base of specific gene protein detection indexes for precision medicine molecular diagnosis of various viruses, pathogenic bacteria, tumors and cancers, etc. Based on the big data prior knowledge base of specific gene protein detection indexes, the nucleic acid protein molecular diagnosis detection results of the swab sample are analyzed by artificial intelligence or / and big data to obtain specific gene protein detection and identification results of various biomedical indexes.

[0085] The wired or wireless communication interface is used to transmit the specific gene protein detection and identification results to the mobile phone of the relevant personnel, or directly report to the cloud big data server or the national epidemic infectious disease, major chronic disease, malignant tumor and cancer health epidemic prevention and health risk monitoring network platform, realize the intelligent medical networking between individuals, families, communities, township clinics and medical units at all levels, health epidemic prevention departments, and predict and warn high-incidence infectious diseases, major chronic diseases, malignant tumors and cancers, etc.

[0086] The display terminal is used to display the specific gene protein detection and identification results of various biomedical indexes.

[0087] Further, the intelligent analysis system 90 also has an artificial intelligence, machine learning and other intelligent analysis function of multi-index joint detection results, further improving the accuracy and reliability of nucleic acid or protein molecule diagnosis results.

[0088] In a preferred embodiment of the present application, the microfluidic chip 10, optical detection system 40, temperature control system 50, heating unit 60, motion control system 80 and intelligent analysis system 90 in the injection tube type sample preparation full integration nucleic acid amplification or protein immune reaction intelligent detection system can be combined for use, forming an independent nucleic acid protein detection intelligent analysis system, which can be compatible with nucleic acid / protein samples obtained by conventional sample preparation methods, and the samples are added to the multi-sample multi-index quantity flexible switching microfluidic chip 10 by manual method, and then the corresponding optical detection system 40, temperature control system 50, heating unit 60, motion control system 80 and / or intelligent analysis system 90 are combined to perform intelligent detection analysis of nucleic acid amplification or protein immune reaction.

[0089] In a preferred embodiment of the present application, as shown in Figure 6 The microprocessor signal storage processing and analysis unit of the injection tube type nucleic acid / protein extraction and purification sample preparation unit 20, reagent storage unit 30, temperature control system 50, heating and shaking unit 70, motion control system 80 and intelligent analysis system 90 can also be used alone to become an independent nucleic acid / protein extraction and purification and concentration sample preparation system to meet the needs of other detection methods requiring nucleic acid proteins.

[0090] In summary, the intelligent detection system provided by the present application can be operated by the user while performing specific gene protein detection of multiple samples, two or more (up to hundreds) of pathogenic bacteria (bacteria, fungi, viruses, microorganisms, model organisms, etc.) infection indicators or tumor and cancer analysis indicators in parallel. The sample reagent consumption of each detection indicator can be reduced from tens of microliters to less than 1 microliter as needed. In addition, the simple valve type flow path isolation technology and multi-sample multi-index quantity flexible switching microfluidic chip provided by the present application effectively improve the universality of the microfluidic chip and realize flexible change and switching of the sample quantity and detection index quantity.

[0091] Each of the embodiments in the specification is described in a progressive manner, and the same or similar parts between the embodiments can be referred to each other. Each of the embodiments focuses on the difference from other embodiments. In the description of the specification, the description of the terms "one preferred embodiment", "further", "specifically", "in this embodiment", and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the embodiments of the specification. In the specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. Furthermore, the person skilled in the art can combine and combine the different embodiments or examples described in the specification and the features of the different embodiments or examples without contradiction.

[0092] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, and not to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A microfluidic chip with flexible switching of multiple sample and index quantities, characterized in that, The microfluidic chip includes: Chip body; The sample inlet channel is provided on the chip body, and the sample inlet channel is provided with a plurality of sample feeding holes; The chip body is provided with at least one exhaust pipe. Each exhaust pipe is connected to the corresponding sample injection pipe through a connecting pipe and a microcavity. The two ends of a microcavity are respectively connected to the sample injection pipe and the exhaust pipe through a connecting pipe to form a detection channel. The number of detection channels is set to multiple. Each exhaust pipe is provided with at least one exhaust hole. An isolation chamber is provided on the sample inlet pipe between adjacent sample inlets, wherein: each isolation chamber is provided with a simple valve, the simple valve adopts a microstructure, the microstructure can deform the isolation chamber pipe by pressurization, thermo-pressurization or ultrasound to achieve flow path blockage and isolation, so as to group the sample inlet pipe and the micro chamber; Alternatively, each of the isolation cavities may optionally contain a simple valve made of an expanding material, which, upon expansion, can achieve flow path blocking and isolation to group the sample inlet pipe and microcavities.

2. The microfluidic chip according to claim 1, characterized in that, The swelling material is a water-swellable material, which includes starch-grafted acrylate polymer crosslinker, acrylamide-acrylate copolymer crosslinker and / or hydrogel resin. When a sample is added to the injection pipe, the water-swellable material expands upon contact with water to achieve flow path blocking and isolation.

3. The microfluidic chip according to claim 1, characterized in that, The expansion material is a heat-expanding material, which includes liquid metal, shape memory alloy, thermally expandable polymer material and / or thermally expandable polymer gel. When the pipeline is heated to a set temperature, the heat-expanding material expands to achieve flow path blockage and isolation.

4. The microfluidic chip according to claim 1, characterized in that, The isolation cavity on the connecting pipe includes: the isolation cavity on the connecting pipe being located at any tangent position to the connecting pipe; the isolation cavity on the connecting pipe being located at a position where it intersects with a portion of the connecting pipe; or the isolation cavity on the connecting pipe being connected to the connecting pipe via a section of pipe.

5. The microfluidic chip according to claim 1, characterized in that, The vent is covered with a waterproof and breathable membrane, and the gas is discharged from the vent through the vent pipe; and / or the sample inlet is covered with a sealing silicone rubber gasket to prevent sample contamination; and / or the sample inlet pipe is straight or curved, with sample inlets at both ends and several sample inlets spaced apart in the middle as needed.

6. An intelligent detection system, characterized in that, Includes the microfluidic chip, injection tube sample preparation unit, reagent storage unit, optical detection system, temperature control system, heating unit, heating and oscillation unit, motion control system and / or intelligent analysis system as described in any one of claims 1 to 5; The injection tube sample preparation unit is configured to lyse the sample and extract, purify, and concentrate nucleic acid proteins. The reagent storage unit is configured to provide reagent storage for sample preparation in the injection tube sample preparation unit and for microfluidic chip reaction. The optical detection system is configured to detect the result of the reaction of the microfluidic chip; The temperature control system is configured to perform constant temperature or variable temperature heating and temperature control on the sample preparation process in the injection tube sample preparation unit, and / or perform constant temperature or cyclic variable temperature heating and temperature control on the microfluidic chip reaction process. The heating and oscillation unit is configured to oscillate and / or heat the injection tube sample preparation unit; The heating unit is configured to heat and / or cool the microfluidic chip; The motion control system is configured to control the sample preparation process in the injection tube sample preparation unit, and / or to control the motion of the microfluidic chip during sample addition and / or reaction. The intelligent analysis system is configured to perform human-machine interactive control of the sample preparation process and / or the reaction process of the microfluidic chip in the injection tube sample preparation unit, and / or to store, analyze, display and / or issue early warnings for the sample reaction detection results acquired by the optical detection system.

7. The intelligent detection system according to claim 6, characterized in that, The injection tube sample preparation unit includes an injection tube, a piston, a filter membrane or filter paper, a sealing ring, a needle, and a protective sleeve. The piston is movably disposed inside the injection tube. The filter membrane or filter paper and the sealing ring are disposed at the bottom of the injection tube. The needle is disposed at the bottom outlet of the injection tube, and the protective sleeve is disposed on the needle.

8. The intelligent detection system according to claim 7, characterized in that, The injection tube is pre-stored with swab preservation elution and / or surface-modified magnetic beads, glass beads, nanomaterials or polymer materials. The surface modification treatment includes anion exchange-type broad-spectrum modification that results in negative charge in acidic environments, and specific group modification based on nucleic acid hybridization or protein immune binding principles that targets the target of interest. This allows for the adsorption of negatively charged nucleic acid proteins in acidic environments, the release of negatively charged nucleic acid proteins in alkaline environments, the adsorption of nucleic acid proteins at low temperatures and / or the release of nucleic acid proteins at high temperatures.

9. The intelligent detection system according to any one of claims 6 to 8, characterized in that, The microfluidic chip, injection tube sample preparation unit, reagent storage unit, optical detection system, temperature control system, heating unit, heating and oscillation unit, motion control system, and intelligent analysis system are all used in combination to achieve fully integrated raw sample input and nucleic acid and protein analysis results output; or, the injection tube sample preparation unit, reagent storage unit, temperature control system, heating and oscillation unit, motion control system, and intelligent analysis system are used in combination to form an independent sample processing system for sample lysis and nucleic acid and protein extraction, purification, and concentration; or, the microfluidic chip, optical detection system, temperature control system, heating unit, motion control system, and intelligent analysis system are used in combination to form an independent intelligent analysis system for nucleic acid and protein detection.

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