Microfluidic chip and centrifugal device for nucleic acid extraction and nucleic acid amplification detection

Abstract

This application relates to the field of nucleic acid detection, disclosing a microfluidic chip and centrifuge device for nucleic acid extraction and amplification detection. The chip includes a base and several sample detection modules arranged around a rotation center of the base. One side of the nucleic acid extraction unit of each sample detection module is connected to a waste liquid channel, and the opposite side is connected to a washing and elution unit and a nucleic acid amplification detection unit. The sample detection module is detachably connected to the base. The washing and elution unit is used to wash and elute the nucleic acid in the sample from the nucleic acid extraction unit under centrifugal force, and the nucleic acid amplification detection unit is used to amplify and detect the eluted sample under centrifugal force. By changing the position of each unit in the sample detection module relative to the rotation center of the base and utilizing centrifugal force to drive reagent flow for nucleic acid extraction and amplification detection, this application's chip does not integrate any pump or valve structure, reducing the complexity, manufacturing difficulty, and cost of the microfluidic chip.

Description

Technical Field

[0001] This application relates to the field of nucleic acid detection, and in particular to a microfluidic chip and centrifugation device for nucleic acid extraction and nucleic acid amplification detection. Background Technology

[0002] Diarrheal diseases are a group of illnesses caused by multiple pathogens and factors, characterized primarily by increased frequency and altered stool consistency. They can lead to severe dehydration, electrolyte imbalance, growth retardation, and even death. Children are the primary affected population, and this is a key disease for prevention and control in my country. Besides the consistently high incidence in clinical cases, viral diarrhea can also cause outbreaks, frequently occurring in childcare facilities and schools. Therefore, rapid and accurate etiological diagnosis is crucial for clinicians to develop appropriate treatment plans and for disease control departments to implement effective measures to control disease outbreaks.

[0003] Methods for detecting the pathogens of viral diarrhea include electron microscopy, virus isolation and culture, immunodiagnostics, and molecular biological diagnostics. The development of microfluidics technology at the beginning of this century has led to the development of nucleic acid detection based on microfluidic chips, which utilizes the basic principles of RT-PCR or isothermal amplification for nucleic acid extraction and amplification. Currently, microfluidic chips integrate numerous micropumps and microvalves, increasing their complexity, manufacturing difficulty, and cost.

[0004] Therefore, how to reduce the complexity, manufacturing difficulty and cost of microfluidic chips is a problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0005] The purpose of this application is to provide a microfluidic chip and centrifugation device for nucleic acid extraction and nucleic acid amplification detection, which reduces the complexity, manufacturing difficulty and cost of microfluidic chips.

[0006] To address the aforementioned technical problems, this application provides a microfluidic chip for nucleic acid extraction and nucleic acid amplification detection, comprising a base and a plurality of sample detection modules arranged around the rotation center of the base;

[0007] The sample detection module includes a nucleic acid extraction unit, a washing and elution unit, a nucleic acid amplification and detection unit, and a waste liquid channel. One side of the nucleic acid extraction unit is connected to the waste liquid channel, and the opposite side of the waste liquid channel is connected to the washing and elution unit and the nucleic acid amplification and detection unit.

[0008] The sample detection module is detachably connected to the base. The washing and elution unit is used to wash and elute the nucleic acid in the sample from the nucleic acid extraction unit under the drive of centrifugal force. The nucleic acid amplification and detection unit is used to amplify and detect the eluted sample under the drive of centrifugal force.

[0009] Preferably, the nucleic acid extraction unit comprises:

[0010] The nucleic acid extraction chamber is used to store magnetic bead reagents;

[0011] A first sample loading well is provided on the nucleic acid extraction chamber for adding sample and lysis buffer into the nucleic acid extraction chamber.

[0012] Preferably, the washing and elution unit includes:

[0013] A cleaning fluid chamber and a second sample dispensing port provided on the cleaning fluid chamber, the second sample dispensing port being used to add cleaning fluid to the cleaning fluid chamber;

[0014] The elution chamber and a third sample feeding port provided on the elution chamber, the third sample feeding port being used to add elution solution to the elution chamber;

[0015] A cleaning solution channel is provided to connect the cleaning solution chamber and the nucleic acid extraction chamber.

[0016] The elution channel is used to connect the elution chamber and the nucleic acid extraction chamber.

[0017] Preferably, the nucleic acid amplification detection unit includes:

[0018] The nucleic acid amplification detection chamber is used to store amplification detection reaction reagents;

[0019] An exhaust port is provided on the nucleic acid amplification detection chamber;

[0020] The sample introduction channel is used to connect the nucleic acid amplification and detection chamber and the nucleic acid extraction chamber.

[0021] Preferably, both the cleaning fluid channel and the elution fluid channel are equipped with capillary valves.

[0022] Preferably, there are multiple cleaning fluid chambers and multiple elution fluid chambers, and the number of cleaning fluid chambers is less than the number of elution fluid chambers;

[0023] Multiple cleaning fluid chambers are arranged sequentially along the rotation center of the base to the edge of the base, and the cleaning fluid channel connects multiple cleaning fluid chambers sequentially. The second sample loading port is located on the cleaning fluid chamber that is furthest from the nucleic acid extraction chamber.

[0024] Multiple elution chambers are arranged sequentially along the rotation center of the base to the edge of the base, and the elution channel is sequentially connected to multiple washing chambers. The third sample loading port is located on the elution chamber that is furthest from the nucleic acid extraction chamber.

[0025] The length of the cleaning fluid channel is less than the length of the elution fluid channel.

[0026] Preferably, a waste liquid collection bottle is connected to the end of the waste liquid channel.

[0027] Preferably, the base has a mounting slot at its rotation center, which is used to mount the microfluidic chip onto the centrifuge device.

[0028] Preferably, the sample detection module is provided with a mounting hole, the base is provided with a mounting post, and the sample detection module and the base are connected to the mounting post through the mounting hole.

[0029] This application also provides a centrifugation device, including the microfluidic chip described above for nucleic acid extraction and nucleic acid amplification detection.

[0030] This application provides a microfluidic chip for nucleic acid extraction and amplification detection, comprising a base and several sample detection modules arranged around the center of rotation of the base. Each sample detection module includes a nucleic acid extraction unit, a washing and elution unit, a nucleic acid amplification detection unit, and a waste liquid channel. One side of the nucleic acid extraction unit is connected to the waste liquid channel, and the opposite side of the waste liquid channel is connected to the washing and elution unit and the nucleic acid amplification detection unit. The sample detection module is detachably connected to the base. The washing and elution unit is used to wash and elute the nucleic acid in the sample from the nucleic acid extraction unit under centrifugal force, and the nucleic acid amplification detection unit is used to amplify and detect the eluted sample under centrifugal force. The sample detection module is detachably connected to the base, and the position of each unit within the sample detection module relative to the center of rotation of the base can be changed. Centrifugal force is used to drive reagent flow for nucleic acid extraction and amplification detection. This chip does not integrate any pump or valve structure; it drives reagent flow through centrifugal force, reducing the complexity, manufacturing difficulty, and cost of the microfluidic chip. Integrating nucleic acid extraction and amplification detection into one unit, nucleic acid extraction and amplification are performed in the same space, eliminating the need for an additional extraction process. The final detection results can be quickly and clearly interpreted by the instrument. Not only does it overcome the shortcomings of traditional nucleic acid testing, such as large workload and long time consumption, but multiple sample detection modules can also simultaneously achieve multi-target detection, enabling the simultaneous detection of multiple viruses.

[0031] The centrifugation device provided in this application includes the microfluidic chip mentioned above for nucleic acid extraction and nucleic acid amplification detection, with the same effect as described above. Attached Figure Description

[0032] To more clearly illustrate the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0033] Figure 1 A structural diagram of a microfluidic chip provided in an embodiment of this application;

[0034] Figure 2 This is a structural diagram of another microfluidic chip provided in an embodiment of this application;

[0035] The attached diagram is labeled as follows: 1 is the base, 2 is the sample detection module, 101 is the mounting slot, 201 is the nucleic acid extraction chamber, 202 is the first sample loading port, 203 is the waste liquid channel, 204 is the cleaning solution chamber, 205 is the second sample loading port, 206 is the cleaning solution channel, 207 is the elution solution chamber, 208 is the third sample loading port, 209 is the elution solution channel, 210 is the nucleic acid amplification detection chamber, 211 is the vent, 212 is the sample injection channel, and 213 is the mounting hole. Detailed Implementation

[0036] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of this application.

[0037] Methods for detecting viral diarrhea pathogens include electron microscopy, virus isolation and culture, immunodiagnostics, and molecular biology diagnostic methods. Electron microscopy allows direct observation of the virus's shape with high resolution, but its sensitivity is low. Virus isolation, culture, and identification are the gold standard for viral pathogen diagnosis, but they are cumbersome, expensive, time-consuming, and have low sensitivity. Commonly used immunodiagnostic methods include enzyme-linked immunosorbent assay (ELISA), immunogold assay, indirect immunofluorescence assay, and latex agglutination assay. These methods are simple and inexpensive, allowing for preliminary screening and detection of samples; however, the sensitivity and specificity of different reagents vary significantly, resulting in relatively poor sensitivity. Molecular biology methods can design primers based on the genome sequences of various viruses and use traditional RT-PCR or real-time quantitative PCR to analyze the amplified DNA products for further viral typing, exhibiting good sensitivity and specificity. However, PCR is a temperature-dependent amplification technique, requiring sophisticated equipment, which hinders its widespread application in point-of-care testing (POCT). Furthermore, PCR only amplifies one-fold of the product per amplification cycle, limiting its application in point-of-care testing (POCT). 9 The process of producing multiple times the product takes a long time, which is not conducive to the need for rapid detection.

[0038] The development of microfluidics at the beginning of this century has made rapid multi-target molecular detection possible. Microfluidic chip technology, relying on modern microfabrication processes, handles or manipulates microfluidics at the micrometer scale. It integrates basic operational units such as sample preparation, reaction, separation, and detection in biological, chemical, and medical analyses onto a single micrometer-scale chip, automating the entire analytical process. This reduces sample and reagent consumption, improves detection sensitivity, shortens reaction time, and lowers average costs. This method overcomes the shortcomings of traditional detection methods and technologies, fundamentally changing and revolutionizing traditional analytical processes and detection methods, and is widely applied to all aspects of traditional detection and analysis. The flexible combination and large-scale integration of multiple unit technologies on a small, controllable platform, characteristic of microfluidic chips, has made them the preferred technology for modern point-of-care testing (POCT) systems. It can be used for microbial pathogen identification, gene mutation detection, genotyping, etc., and is currently an effective method for rapid multi-target detection of pathogenic microorganisms.

[0039] The core of this application is to provide a microfluidic chip and centrifugation device for nucleic acid extraction and nucleic acid amplification detection.

[0040] To enable those skilled in the art to better understand the present application, the present application will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0041] like Figure 1As shown, this application embodiment provides a microfluidic chip for nucleic acid extraction and nucleic acid amplification detection, including a base 1 and a plurality of sample detection modules 2 arranged around the rotation center of the base 1; the sample detection module 2 includes a nucleic acid extraction unit, a washing and elution unit, a nucleic acid amplification detection unit, and a waste liquid channel 203. One side of the nucleic acid extraction unit is connected to the waste liquid channel 203, and the opposite side of the waste liquid channel 203 is connected to the washing and elution unit and the nucleic acid amplification detection unit; the sample detection module 2 is detachably connected to the base 1. The washing and elution unit is used to wash and elute the nucleic acid in the sample of the nucleic acid extraction unit under the drive of centrifugal force, and the nucleic acid amplification detection unit is used to amplify and detect the eluted sample under the drive of centrifugal force.

[0042] The shape of the base 1 is not specifically limited in this embodiment of the application, such as Figure 1 As shown, the base 1 can be circular. The specific method by which the base 1 and the sample detection module 2 are detachably connected is not limited; for example, the sample detection module 2 may have a mounting hole 213, and the base 1 may have a mounting post, with the sample detection module 2 and base 1 connected to the mounting post through the mounting hole 213. The sample detection module 2 can be disposable, while the base 1 can be reused. This application embodiment does not specify the number of sample detection modules 2; the sample detection modules 2 are arranged around the rotation center of the base 1, such as... Figure 1 As shown, the base is circular, and the four sample detection modules 2 are identical in shape and size. Since the sample detection modules 2 are detachably connected to the base 1, the relative positions of the cleaning and elution unit, nucleic acid amplification and detection unit, and waste liquid channel 203 within the sample detection module 2 with respect to the rotation center of the base 1 can be changed. When cleaning and eluting samples, the sample detection modules 2 are installed as follows: Figure 1 As shown, the waste liquid channel 213 is positioned away from the rotation center of the base 1, so that the washing and elution unit is close to the rotation center of the base 1. Driven by centrifugal force, the washing liquid and elution liquid flow into the nucleic acid extraction unit for nucleic acid washing and elution. The waste liquid after washing and elution is discharged through the waste liquid channel 213. When performing isothermal amplification detection on the sample, when installing the sample detection module 2, as shown... Figure 2 As shown, the waste liquid channel 213 is placed close to the rotation center of the base 1 so that the nucleic acid amplification detection unit is far away from the rotation center of the base 1. Under the drive of centrifugal force, the nucleic acid sample after washing and elution in the nucleic acid extraction unit flows into the nucleic acid amplification detection unit for amplification and detection.

[0043] This application provides a microfluidic chip for nucleic acid extraction and amplification detection, comprising a base and several sample detection modules arranged around the center of rotation of the base. Each sample detection module includes a nucleic acid extraction unit, a washing and elution unit, a nucleic acid amplification detection unit, and a waste liquid channel. One side of the nucleic acid extraction unit is connected to the waste liquid channel, and the opposite side is connected to the washing and elution unit and the nucleic acid amplification detection unit. The sample detection module is detachably connected to the base. The washing and elution unit is used to wash and elute the nucleic acid in the sample from the nucleic acid extraction unit under centrifugal force, and the nucleic acid amplification detection unit is used to amplify and detect the eluted sample under centrifugal force. By changing the position of each unit in the sample detection module relative to the center of rotation of the base and using centrifugal force to drive reagent flow for nucleic acid extraction and amplification detection, this application's chip does not integrate any pump or valve structure, reducing the complexity, manufacturing difficulty, and cost of the microfluidic chip. Integrating nucleic acid extraction and amplification detection into one unit, nucleic acid extraction and amplification are performed in the same space, eliminating the need for an additional extraction process. The final detection results can be quickly and clearly interpreted using a photomultiplier tube. Not only can it overcome the shortcomings of traditional nucleic acid testing, such as large workload and long time consumption, but multiple sample detection modules can also realize multi-target detection at the same time, and can detect multiple viruses simultaneously.

[0044] Based on the above embodiments, the nucleic acid extraction unit in this application includes: a nucleic acid extraction chamber 201 for storing magnetic bead reagents; and a first sample application well 202 disposed on the nucleic acid extraction chamber 201 for adding samples and lysis buffer into the nucleic acid extraction chamber 201. Specifically, the nucleic acid extraction chamber 201 contains magnetic bead reagents obtained through vacuum freeze-drying.

[0045] Based on the above embodiments, the washing and elution unit of this application includes: a washing liquid chamber 204 and a second sample feeding port 205 disposed on the washing liquid chamber 204, the second sample feeding port 205 being used to add washing liquid to the washing liquid chamber 204; an elution liquid chamber 207 and a third sample feeding port 208 disposed on the elution liquid chamber 207, the third sample feeding port 208 being used to add elution liquid to the elution liquid chamber 207; a washing liquid channel 206 being used to connect the washing liquid chamber 204 and the nucleic acid extraction chamber 201; and an elution liquid channel 209 being used to connect the elution liquid chamber 207 and the nucleic acid extraction chamber 201.

[0046] Based on the above embodiments, the nucleic acid amplification detection unit of this application includes: a nucleic acid amplification detection chamber 210 for storing amplification detection reaction reagents; an exhaust port 211 disposed on the nucleic acid amplification detection chamber 210; and a sample inlet channel 212 for connecting the nucleic acid amplification detection chamber 210 and the nucleic acid extraction chamber 201. The nucleic acid amplification detection chamber 210 contains nucleic acid amplification reagents and detection reagents obtained through vacuum freeze-drying. The nucleic acid amplification process temperature is 60-65℃, eliminating the need for sophisticated temperature control equipment and making it suitable for on-site sample point diagnostic testing.

[0047] like Figure 1 As shown, one side of the nucleic acid extraction chamber 201 is connected to the washing solution chamber 204 via the washing solution channel 206, and to the elution solution chamber 207 via the elution solution channel 209. It is also connected to the nucleic acid amplification and detection chamber 210 via the sample injection channel 212. The washing solution chamber 204, elution solution chamber 207, and nucleic acid amplification and detection chamber 210 are located on the same side of the nucleic acid extraction chamber 201, while the waste liquid channel 203 is located on the opposite side. If the base 1 is circular, the washing solution channel 206, elution solution channel 209, waste liquid channel 203, and sample injection channel 212 should coincide with the radius of the circular base, which is more conducive to the centrifugal force driving the liquid.

[0048] Based on the above embodiments, both the cleaning fluid channel and the elution fluid channel in this application embodiment are equipped with capillary valves.

[0049] During washing and elution, the washing solution and elution solution flow to the nucleic acid extraction chamber 201 under high-speed centrifugation to wash and elute the sample in the nucleic acid extraction chamber 201. During nucleic acid isothermal amplification detection, under low-speed centrifugation, the centrifugal force is less than the capillary force, making it difficult for the eluted sample to enter the washing solution chamber 204 and the elution solution chamber 207, and it directly enters the nucleic acid amplification detection chamber 210.

[0050] Based on the above embodiments, there are multiple washing liquid chambers 204 and multiple elution liquid chambers 207, and the number of washing liquid chambers 204 is less than the number of elution liquid chambers 207; the multiple washing liquid chambers 204 are arranged sequentially along the direction from the rotation center of the base 1 to the edge of the base 1, and the washing liquid channel 206 is connected to the multiple washing liquid chambers 204 in sequence, and the second sample loading port 205 is located on the washing liquid chamber 204 that is farthest from the nucleic acid extraction chamber 201; the multiple elution liquid chambers 207 are arranged sequentially along the direction from the rotation center of the base 1 to the edge of the base 1, and the elution liquid channel 209 is connected to the multiple washing liquid chambers 207 in sequence, and the third sample loading port 208 is located on the elution liquid chamber 209 that is farthest from the nucleic acid extraction chamber 201; wherein, the length of the washing liquid channel 206 is less than the length of the elution liquid channel 109.

[0051] The embodiments of this application do not limit the number of cleaning fluid chambers 206 and elution fluid chambers 207. Of course, multiple thorough cleanings can be achieved by increasing the number or volume of the cleaning fluid chambers. Figure 1 As shown, the washing solution chamber 204 may include a primary washing solution chamber and a secondary washing solution chamber. The primary washing solution chamber has a second sample application port 205. That is, the distance between the primary washing solution chamber and the nucleic acid extraction chamber 201 is greater than the distance between the secondary washing solution chamber and the nucleic acid extraction chamber 201. The elution chamber 207 may include a primary elution chamber, a secondary elution chamber, a tertiary elution chamber, and a quaternary elution chamber, arranged sequentially from farthest from the nucleic acid extraction chamber 201. A third sample application port 208 is located in the primary elution chamber. In this embodiment, the number of washing solution chambers 204 is less than the number of elution chambers 207, and the length of the washing solution channel 206 is less than the length of the elution channel 209. This is all to ensure that the washing solution flows into the nucleic acid extraction chamber 201 before the elution solution, so as to wash the sample first, and then elute the sample. Of course, there can be multiple nucleic acid amplification detection chambers 210. Each nucleic acid amplification detection chamber 210 is equipped with an exhaust port 211. Each nucleic acid amplification detection chamber 210 can store different viral reaction solutions, containing primers and probes, to detect different viruses. Specifically, the nucleic acid extraction chamber 201 has a volume of 1300 μL, the washing solution chamber 204 has a volume of 500 μL, the elution solution chamber 207 has a volume of 200 μL, and the nucleic acid amplification detection chamber 210 has a volume of 25 μL.

[0052] Based on the above embodiments, the tail end of the waste liquid channel 203 in this application embodiment is connected to a waste liquid collection bottle.

[0053] The diameter of the waste liquid channel 203 is three times that of other channels, which facilitates the rapid discharge of waste liquid. An external waste liquid collection bottle is connected, and the waste liquid is directly discharged into the waste liquid collection bottle through the waste liquid channel 203 to prevent the waste liquid from entering the nucleic acid amplification detection chamber 210 in reverse after the installation direction of the sample detection module 2 is changed.

[0054] Based on the above embodiments, the base 1 of this application embodiment is provided with a mounting slot 101 at the rotation center, and the mounting slot 101 is used to install the microfluidic chip on the centrifuge device.

[0055] Based on this, the detection process is described below. The sample detection module 2 is positioned on the base 1 with the centrifugal force directed towards the waste liquid channel 203, i.e., the waste liquid channel 203 is away from the rotation center of the base 1. 400 μL of the processed sample and 850 μL of lysis buffer are sequentially injected into the nucleic acid extraction chamber through the first sample loading port 202. 500 μL of washing buffer is injected into the primary washing buffer chamber through the second sample loading port 205. 200 μL of elution buffer is injected into the primary elution buffer chamber through the third sample loading port 208. Virus lysis releases nucleic acid, which is adsorbed onto magnetic beads. After a 10-minute reaction, the magnetic beads are fixed by magnet adsorption, and the mixture is centrifuged at 1500 rpm for 20 seconds. Under the combined effects of centrifugal force and gravity, the waste liquid is discharged into the waste liquid collection bottle. Simultaneously, the washing buffer enters the secondary washing buffer chamber from the primary washing buffer chamber, and the elution buffer enters the secondary elution buffer chamber from the primary elution buffer chamber. Continue centrifugation at 1500 rpm for 20 seconds. The washing solution enters the nucleic acid extraction chamber 201 from the secondary washing solution chamber, while the elution solution enters the tertiary elution chamber from the secondary elution chamber. Centrifugation at 1500 rpm for 20 seconds drives the washing solution and other reagents in the nucleic acid extraction chamber 201 to drain into the waste collection bottle. Simultaneously, the elution solution enters the quaternary elution chamber from the tertiary elution chamber. Centrifugation at 1500 rpm for 20 seconds drives the elution solution from the quaternary elution chamber into the nucleic acid extraction chamber, eluting the nucleic acid adsorbed on the magnetic beads. The elution time is 5 minutes. After elution, rotate the sample detection module 2 180° and fix it to the base 1, so that the centrifugal force is directed towards the nucleic acid amplification detection chamber 210, i.e., the waste channel 203 is close to the rotation center of the base 1. Centrifugation at 500 rpm for 10 seconds drives the eluted nucleic acid sample from the nucleic acid extraction chamber 201 to the five nucleic acid amplification detection chambers 210. The viral load in the sample is quantified by isothermal amplification detection.

[0056] Finally, this application also provides a centrifugation device, including the microfluidic chip mentioned above for nucleic acid extraction and nucleic acid amplification detection. Since the microfluidic chip has been described in detail in the above embodiments, it will not be repeated in this application.

[0057] This application provides a centrifugation device including the aforementioned microfluidic chip for nucleic acid extraction and nucleic acid amplification detection. The microfluidic chip includes a base and several sample detection modules arranged around the center of rotation of the base. Each sample detection module includes a nucleic acid extraction unit, a washing and elution unit, a nucleic acid amplification detection unit, and a waste liquid channel. One side of the nucleic acid extraction unit is connected to the waste liquid channel, and the opposite side of the waste liquid channel is connected to the washing and elution unit and the nucleic acid amplification detection unit. The sample detection module is detachably connected to the base. The washing and elution unit is used to wash and elute the nucleic acid in the sample from the nucleic acid extraction unit under centrifugal force, and the nucleic acid amplification detection unit is used to amplify and detect the eluted sample under centrifugal force. By changing the position of each unit in the sample detection module relative to the center of rotation of the base and using centrifugal force to drive reagent flow for nucleic acid extraction and nucleic acid amplification detection, the chip of this application does not integrate any pump or valve structure. By using centrifugal force to drive reagent flow, the complexity, manufacturing difficulty, and cost of the microfluidic chip are reduced. Integrating nucleic acid extraction and amplification detection into one unit, nucleic acid extraction and amplification are performed in the same space, eliminating the need for an additional extraction process. The final detection results can be quickly and clearly interpreted by the instrument. Not only does it overcome the shortcomings of traditional nucleic acid testing, such as large workload and long time consumption, but multiple sample detection modules can also simultaneously achieve multi-target detection, enabling the simultaneous detection of multiple viruses.

[0058] The foregoing has provided a detailed description of a microfluidic chip and centrifugation device for nucleic acid extraction and amplification detection provided in this application. The various embodiments in the specification are described in a progressive manner, with each embodiment focusing on its differences from other embodiments. Similar or identical parts between embodiments can be referred to interchangeably. It should be noted that those skilled in the art can make various improvements and modifications to this application without departing from the principles thereof, and these improvements and modifications also fall within the protection scope of the claims of this application.

[0059] It should also be noted that, in this specification, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

Claims

1. A microfluidic chip for nucleic acid extraction and nucleic acid amplification detection, characterized in that, It includes a base and several sample detection modules arranged around the rotation center of the base; The sample detection module includes a nucleic acid extraction unit, a washing and elution unit, a nucleic acid amplification and detection unit, and a waste liquid channel. One side of the nucleic acid extraction unit is connected to the waste liquid channel, and the opposite side of the waste liquid channel is connected to the washing and elution unit and the nucleic acid amplification and detection unit. The sample detection module is detachably connected to the base. The washing and elution unit is used to wash and elute the nucleic acid in the sample from the nucleic acid extraction unit under the drive of centrifugal force. The nucleic acid amplification and detection unit is used to amplify and detect the eluted sample under the drive of centrifugal force. The nucleic acid extraction unit includes: The nucleic acid extraction chamber is used to store magnetic bead reagents; A first sample loading well is provided on the nucleic acid extraction cavity for adding sample and lysis buffer into the nucleic acid extraction cavity; The washing and elution unit includes: A cleaning fluid chamber and a second sample dispensing port provided on the cleaning fluid chamber, the second sample dispensing port being used to add cleaning fluid to the cleaning fluid chamber; The elution chamber and a third sample feeding port provided on the elution chamber, the third sample feeding port being used to add elution solution to the elution chamber; A cleaning solution channel is provided to connect the cleaning solution chamber and the nucleic acid extraction chamber. An elution channel is provided to connect the elution chamber and the nucleic acid extraction chamber. The nucleic acid amplification and detection unit includes: The nucleic acid amplification detection chamber is used to store amplification detection reaction reagents; An exhaust port is provided on the nucleic acid amplification detection chamber; A sample inlet channel is used to connect the nucleic acid amplification and detection chamber and the nucleic acid extraction chamber; There are multiple cleaning fluid chambers and multiple elution fluid chambers, and the number of cleaning fluid chambers is less than the number of elution fluid chambers; Multiple cleaning fluid chambers are arranged sequentially along the rotation center of the base to the edge of the base, and the cleaning fluid channel connects multiple cleaning fluid chambers sequentially. The second sample loading port is located on the cleaning fluid chamber that is furthest from the nucleic acid extraction chamber. Multiple elution chambers are arranged sequentially along the rotation center of the base to the edge of the base, and the elution channel is sequentially connected to multiple washing chambers. The third sample loading port is located on the elution chamber that is furthest from the nucleic acid extraction chamber. Wherein, the length of the cleaning fluid channel is less than the length of the elution fluid channel; The base is circular, and the cleaning solution channel, elution solution channel, waste liquid channel, and sample injection channel should coincide with the radius of the circular base.

2. The microfluidic chip for nucleic acid extraction and nucleic acid amplification detection according to claim 1, characterized in that, Both the cleaning fluid channel and the elution fluid channel are equipped with capillary valves.

3. The microfluidic chip for nucleic acid extraction and nucleic acid amplification detection according to claim 1, characterized in that, The waste liquid channel is connected to a waste liquid collection bottle at its end.

4. The microfluidic chip for nucleic acid extraction and nucleic acid amplification detection according to claim 1, characterized in that, The base has a mounting slot at its rotation center, which is used to mount the microfluidic chip onto the centrifuge device.

5. The microfluidic chip for nucleic acid extraction and nucleic acid amplification detection according to claim 1, characterized in that, The sample detection module is provided with a mounting hole, and the base is provided with a mounting post. The sample detection module and the base are connected to the mounting post through the mounting hole.

6. A centrifuge device, characterized in that, Includes the microfluidic chip for nucleic acid extraction and nucleic acid amplification detection as described in any one of claims 1 to 5.

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