Virus and bacterium typing detection device

Abstract

The utility model discloses a virus and bacterium typing detection device. Comprising an injector, a nucleic acid extraction assembly, a microfluidic amplification detection assembly, a box cover and a box body, and the box cover is mounted on the box body; the injector is arranged in the box body; the nucleic acid extraction assembly and the microfluidic amplification detection assembly are both mounted in the box body. Wherein the micro-fluidic amplification detection assembly comprises a micro-fluidic chip, a heating sheet, a sample adding hole, a virus reaction hole and a bacterium reaction hole; a sample adding hole, a virus reaction hole and a bacteria reaction hole are formed in the micro-fluidic chip, the sample adding hole is respectively communicated with the virus reaction hole and the bacteria reaction hole, and a heating sheet is arranged below the micro-fluidic chip. According to the device, nucleic acid extraction and detection can be completed through simple operation, the nucleic acid amplification reaction process can be completed only through one constant-temperature heating sheet, and meanwhile the device has the advantages of being easy to operate, high in specificity, high in sensitivity, high in flux and the like.

Description

Technical Field

[0001] The utility model relates to a pathogen detection device in the field of biological detection, and particularly to a device for detecting virus and bacteria typing. Background Art

[0002] Nucleic acid extraction is a commonly used technical method in modern molecular biology. Currently, the commonly used methods on the market include phenol-chloroform extraction method, purification method based on centrifugal column kits, and purification method based on magnetic beads. However, these methods have their respective defects.

[0003] The phenol-chloroform extraction method has a relatively low cost, but it will use organic substances such as phenol and chloroform that are toxic to the human body.

[0004] The purification method based on centrifugal column kits is the most widely used method in kits. However, this method uses a special silicon matrix material to bind DNA in an environment of high salt and low pH, while the remaining proteins and RNAs are not bound. After washing, a low-salt and high-pH eluent is used to break the salt bridge between DNA and silicon to elute DNA from the silicon matrix material. This method relies on large-scale instruments such as high-speed centrifuges and has a relatively high cost.

[0005] The magnetic bead method utilizes the principle that the active groups on magnetic particles can bind to and dissociate from nucleic acids under certain conditions. After mixing magnetic beads with the target sample for a period of time, the magnetic particles are aggregated and adsorbed by using a magnetic field, so that the magnetic particles and the liquid are separated. After washing the impurities on the magnetic beads, the nucleic acids on the magnetic beads are eluted with an eluent to obtain the required DNA. This method is simple to operate, but the disadvantages are relatively high cost and most of the magnetic beads sold on the market are polydisperse, resulting in low nucleic acid concentration and many impurities in the extracted nucleic acids, which is not conducive to subsequent detection.

[0006] In addition, the purification method based on magnetic beads requires large-scale facilities such as nucleic acid extractors. At the same time, polymerase chain reaction (PCR) is required for detection after nucleic acid extraction, which relies on large-scale PCR instruments. Most of the application sites are professional testing institutions such as laboratories and hospitals. The experimental conditions are relatively simple in places such as homes, schools, and stations, and it is relatively difficult to maintain this system. Moreover, during the epidemic period, people all go to the hospital for nucleic acid testing, resulting in an increasing possibility of cross-infection among patients.

[0007] At the same time, existing detection methods or biosensors often focus on the detection of a single pathogen, and there are few reports on the ability to simultaneously quantify and detect multiple pathogens in the same sample. People cannot immediately and accurately determine which type of disease they have in the early stage of illness. Therefore, it is particularly important to develop a simple, efficient, sensitive, high-throughput, and highly accurate nucleic acid detection method. Content of the Utility Model

[0008] To overcome the deficiencies of the prior art, the present utility model provides a device for virus and bacteria typing detection, which can complete nucleic acid extraction and detection through simple operations, without the need for special instrument equipment. Only a constant temperature heating sheet is required to complete the nucleic acid amplification reaction process. By visually observing the color change, it is possible to determine the bacteria or virus carried in the sample, and intuitively judge the disease condition of the human body. At the same time, the device is easy to operate, has the characteristics of strong specificity, high sensitivity, high throughput, etc., and is suitable for use in environments without laboratory conditions such as families, farms, ports, and primary community clinics. It can be used in remote and poverty-stricken areas with scarce medical resources. The utility model can greatly improve the integration and portability of virus detection and bacteria detection on the current market.

[0009] The technical solution of the present utility model is as follows:

[0010] The present utility model includes a box cover and a box body, and the box cover is installed on the box body; it also includes a syringe, which is installed in the box body; it further includes a nucleic acid extraction component, which is installed in the box body; and a microfluidic amplification detection component, which is also installed in the box body.

[0011] Further, the nucleic acid extraction component includes a plurality of solution tubes, and all the plurality of solution tubes are placed in the box body. The solution tubes include a lysis solution tube, a first washing solution tube, a second washing solution tube, an elution solution tube, a collection tube, a waste liquid tube, a PBS tube, a primer tube, a dye tube, and a ddH 2 O tube.

[0012] Further, among the solution tubes, the lysis solution tube, the first washing solution tube, the second washing solution tube, the elution solution tube, the collection tube, and the waste liquid tube are arranged in a row in sequence, and the PBS tube, the primer tube, the dye tube, and the ddH 2 O tube are arranged in another row in sequence.

[0013] Further, the bottom of the elution solution tube is a pointed bottom protruding downward.

[0014] Further, the tube bodies of the lysis solution tube, the first washing solution tube, the second washing solution tube, the PBS tube, the primer tube, the dye tube, and the ddH 2 O tube are all flat bottoms.

[0015] Further, the microfluidic amplification detection component includes a microfluidic chip, a heating sheet, a sample addition hole, a virus reaction hole, and a bacteria reaction hole; the microfluidic chip is provided with a sample addition hole, a virus reaction hole, and a bacteria reaction hole. The sample addition hole is respectively communicated with the virus reaction hole and the bacteria reaction hole, and a heating sheet is installed under the microfluidic chip.

[0016] Furthermore, the virus reaction wells include one or more of influenza B virus reaction wells, respiratory syncytial virus reaction wells, human adenovirus reaction wells, influenza A (H1N1) virus reaction wells, and influenza A (H7N9) virus reaction wells.

[0017] Furthermore, the bacterial reaction wells include one or more of Staphylococcus aureus reaction wells, Escherichia coli reaction wells, Streptococcus pneumoniae reaction wells, Klebsiella pneumoniae reaction wells, and Pseudomonas aeruginosa reaction wells.

[0018] Furthermore, the heating plate is a disposable heating plate and can maintain a constant temperature of 65 degrees Celsius for 30 minutes.

[0019] Furthermore, a groove is provided inside the box cover, and the barrel of the syringe can be embedded in the groove.

[0020] Furthermore, a film for sealing the corresponding solution tube is provided on the top of each solution tube.

[0021] Furthermore, the pipe openings of the solution pipes are all equipped with pipe covers that are threadedly connected.

[0022] Furthermore, the microfluidic chip has a snap-on design to protect the solution from contacting the outside world.

[0023] Furthermore, a film for sealing the pipeline is arranged on the top of the microfluidic chip.

[0024] Furthermore, a groove matching a 1 ml syringe is provided inside the cover of the device, so that the syringe can be placed in the card slot when not in use to avoid accidental injury to others and contamination of the needle.

[0025] The solution tube includes the following two sealing methods:

[0026] The first type: the top of each solution tube is provided with a film for sealing the corresponding solution tube, which can be a plastic film or an aluminum foil.

[0027] The second type: The outlets of the solution tubes are all equipped with threaded tube caps.

[0028] Microfluidic chips include the following two sealing methods:

[0029] The first type: A thin film is set on the top of the microfluidic chip to seal all the channels.

[0030] The second type: The microfluidic chip has a snap-on design that protects the solution from contacting the outside world.

[0031] The utility model has the following beneficial effects:

[0032] 1. The device can detect the nucleic acids of 10 common pathogens through a microfluidic chip without the need for large equipment, and the detection can be completed at home, reducing the possibility of cross-infection among patients caused by people going to the hospital for nucleic acid testing during the prevalence of epidemic diseases.

[0033] 2. The device uses a nucleic acid adsorption membrane and its supporting reagents to extract nucleic acids from samples, which has the characteristics of simplicity, high efficiency, non-toxicity, etc. compared with the nucleic acid extraction methods on the market.

[0034] 3. The nucleic acid amplification technology of the device adopts loop-mediated isothermal amplification technology in the microfluidic chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 It is a three-dimensional axonometric view of the device of the present utility model.

[0036] Figure 2 It is a top view of the device of the present utility model.

[0037] Figure 3 It is a schematic diagram of the positions of the reaction holes on the microfluidic chip.

[0038] Figure 4 It is a schematic diagram of the operation of the device of the present utility model Figure 1 .

[0039] Figure 5 It is a schematic diagram of the operation of the device of the present utility model Figure 2 .

[0040] In the figure: 1. Needle, 2. Converter, 3. Rubber ring, 4. Nucleic acid adsorption membrane, 5. Barrel body, 6. Pusher, 7. Microfluidic chip, 8. Heating sheet, 9. Switch buckle, 10. Groove, 11. Box cover, 12. Box body, 111. Lysis solution tube, 112. First washing solution tube, 113. Second washing solution tube, 114. Elution solution tube, 115. Collection tube, 116. Waste liquid tube, 117. PBS tube, 118. Primer tube, 119. Dye tube, 120. ddH 2 O tube, 221. Sampling hole, 222. Reaction hole for influenza B virus, 223. Reaction hole for respiratory syncytial virus, 224. Reaction hole for human adenovirus, 225. Reaction hole for influenza A (H1N1) virus, 226. Reaction hole for influenza A (H7N9) virus, 227. Reaction hole for Staphylococcus aureus, 228. Reaction hole for Escherichia coli, 229. Reaction hole for Streptococcus pneumoniae, 230. Reaction hole for Klebsiella pneumoniae, 231. Reaction hole for Pseudomonas aeruginosa. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0041] The present utility model will be further described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present utility model, and are not used to limit the present utility model.

[0042] As Figure 1 , Figure 2 , Figure 4 and Figure 5 shown, the present utility model includes a syringe, a nucleic acid extraction component, a microfluidic amplification detection component, a box cover 11 and a box body 12, and the box cover 11 is installed on the box body 12; the syringe is installed in the box body 12, specifically installed inside or on the side of the box body 12; the nucleic acid extraction component is installed inside the box body 12; the microfluidic amplification detection component is also installed inside the box body 12.

[0043] One side of the box cover 11 is hinged to the upper part of the box body 12, so that the box cover 11 is arranged on the box body 12 after being opened. A switch buckle 9 is arranged on the other side of the box cover 11, and a protrusion is arranged on the box body 12. The switch buckle 9 cooperates with the protrusion, so that the box cover 11 is fixedly installed on the box body 12. A groove 10 is opened inside the box cover 11, and the barrel 5 of the syringe can be embedded in the groove 10.

[0044] The syringe includes a needle 1, a converter 2, a rubber ring 3, a nucleic acid adsorption membrane 4, a barrel 5 and a push rod 6; the needle 1 is installed at one end of the barrel 5 through the converter 2, a nucleic acid adsorption membrane 4 is arranged on the inner wall of the barrel 5, a push rod 6 is installed inside the barrel 5, a rubber ring 3 is installed at one end of the push rod 6, and one end of the nucleic acid adsorption membrane 4 close to the needle 1 is arranged between the chamber between the rubber ring 3 and the bottom of the barrel 5, that is, one end of the nucleic acid adsorption membrane 4 close to the needle 1 is pressed against the bottom of the barrel 5 by the rubber ring 3.

[0045] The nucleic acid extraction component includes a plurality of solution tubes, and the plurality of solution tubes are all placed inside the box body 12. The solution tubes include a lysis solution tube 111, a first washing solution tube 112, a second washing solution tube 113, an elution solution tube 114, a collection tube 115, a waste liquid tube 116, a PBS tube 117, a primer tube 118, a dye tube 119 and a ddH 2 O tube 120.

[0046] Among the solution tubes, the lysis solution tube 111, the first washing solution tube 112, the second washing solution tube 113, the elution solution tube 114, the collection tube 115 and the waste liquid tube 116 are arranged in a row in sequence, and the PBS tube 117, the primer tube 118, the dye tube 119 and the ddH 2 O tube 120 are arranged in another row in sequence.

[0047] The bottom of the eluent tube 114 is a pointed bottom protruding downward, so that the needle 1 can fully suck away the solution in the eluent tube 114. The lysis solution tube 111, the first washing solution tube 112, the second washing solution tube 113, the PBS tube 117, the primer tube 118, the dye tube 119, and the ddH 2 The tube bodies of the O tube 120 are all flat bottoms, which is beneficial to prompt the user that there is no need to suck away the solution in the corresponding steps of the solution tube.

[0048] The microfluidic amplification detection component includes a microfluidic chip 7, a heating sheet 8, a sample addition hole 221, a viral reaction hole, and a bacterial reaction hole; the microfluidic chip 7 is provided with a sample addition hole 221, a viral reaction hole, and a bacterial reaction hole. The sample addition hole 221 is respectively communicated with the viral reaction hole and the bacterial reaction hole, and the viral reaction hole and the bacterial reaction hole are not communicated with each other. A heating sheet 8 is installed under the microfluidic chip 7 for heating the microfluidic chip 7.

[0049] As Figure 3 shown, the viral reaction holes include one or more of the influenza B virus reaction hole 222, the respiratory syncytial virus reaction hole 223, the human adenovirus reaction hole 224, the influenza A H1N1 virus reaction hole 225, and the influenza A H7N9 virus reaction hole 226. The bacterial reaction holes include one or more of the Staphylococcus aureus reaction hole 227, the Escherichia coli reaction hole 228, the Streptococcus pneumoniae reaction hole 229, the Klebsiella pneumoniae reaction hole 230, and the Pseudomonas aeruginosa reaction hole 231.

[0050] The heating sheet 8 is a disposable heating sheet and can maintain a constant temperature of 65 degrees Celsius for 30 minutes.

[0051] A film for sealing the corresponding solution tube is provided at the top of each solution tube.

[0052] A tube cap with a threaded connection is provided at the mouth of each solution tube.

[0053] The microfluidic chip 7 has a snap design to protect the solution from contacting the outside.

[0054] A film for sealing the pipeline is provided at the top of the microfluidic chip 7.

[0055] A groove 10 matching with a 1 ml syringe is provided inside the cover body of the device, so that the syringe can be placed at the card slot when not in use, so as to avoid hurting others and contaminating the needle.

[0056] The solution tube includes the following two sealing methods:

[0057] The first one: A film for sealing the corresponding solution tube is provided at the top of each solution tube. This film can be a plastic film or an aluminum foil.

[0058] The second one: A tube cap with a threaded connection is provided at the mouth of each solution tube.

[0059] The microfluidic chip includes the following two sealing methods:

[0060] The first one: A film for sealing all pipelines is provided on the top of the microfluidic chip.

[0061] The second one: The microfluidic chip has a snap-fastener design to protect the solution from contacting the outside.

[0062] When in use, this device is simpler than the phenol-chloroform extraction method, the centrifugal column-based purification method, and the magnetic bead-based purification method. It can directly extract and detect nucleic acids from samples on-site without large equipment such as centrifuges. The extracted sample nucleic acids are amplified in the microfluidic chip using loop-mediated isothermal amplification technology and then detected. There are a total of 10 reaction holes on the microfluidic chip, including 5 for viruses and 5 for bacteria.

[0063] The usage method of the integrated virus and bacteria typing detection device provided by the present utility model is as follows:

[0064] First, insert the needle 1 into the bottom of the lysis solution tube 111, add the collected substance to be detected into the lysis solution, and press and lift the push rod 6 for 8 - 10 times to enable the nucleic acid adsorption membrane 4 to fully adsorb nucleic acids; then press the push rod to the bottom, insert the needle of the transfer syringe into the bottom of the first washing tube 112, and press and lift the push rod 6 for 5 - 8 times to wash away some impurities on the nucleic acid adsorption membrane 4; then press the push rod to the bottom again, insert the needle of the transfer syringe into the bottom of the second washing tube 113, and press and lift the push rod 6 for 5 - 8 times to wash away another part of the impurities on the nucleic acid adsorption membrane, then press to the bottom, and transfer the 1 needle to the elution solution tube 114, slowly press and release the push rod 6 for 8 - 10 times to desorb the nucleic acids from the nucleic acid adsorption membrane 4 and transfer the liquid in the tube to the collection tube 115. After that, add the collected liquid into the PBS solution 117, press and lift the push rod 6 for 5 - 8 times, and then add the liquid into the primer tube 118, the dye tube 119, ddH 2 O tube 120. Finally, suck all the liquid into the syringe and inject it into the loading hole 221 at a uniform speed, and then it will flow into the reaction holes of the 10 channels. If the solution contains nucleic acids corresponding to the pathogens in a certain reaction hole, the color of this reaction hole will change from purple to sky blue, enabling people to directly visually observe the results through the color change.

[0065] Finally, it should be noted that the above embodiments and descriptions are only used to illustrate the technical solutions of the present utility model rather than to limit them. Those of ordinary skill in the art should understand that the technical solutions of the present utility model can be modified or equivalently replaced. Without departing from the spirit and scope of the disclosure of the technical solutions of the present utility model, they should all be covered within the protection scope of the claims of the present utility model.

Claims

1. A device for virus and bacteria typing detection, characterized in that: The invention comprises a box cover (11) and a box body (12), wherein the box cover (11) is mounted on the box body (12); a syringe is mounted in the box body (12); a nucleic acid extraction component is mounted in the box body (12); and a microfluidic amplification detection component is also mounted in the box body (12).

2. A device for virus and bacteria typing detection according to claim 1, characterized in that: The nucleic acid extraction component comprises a plurality of solution tubes, all of which are placed in a box body (12), and the solution tubes comprise a lysis solution tube (111), a first washing solution tube (112), a second washing solution tube (113), an elution solution tube (114), a collection tube (115), a waste solution tube (116), a PBS tube (117), a primer tube (118), a dye tube (119) and a ddH2O tube (120).

3. A device for virus and bacteria typing detection according to claim 2, characterized in that: In the solution tubes, the lysate tube (111), the first washing solution tube (112), the second washing solution tube (113), the elution solution tube (114), the collection tube (115) and the waste liquid tube (116) are sequentially arranged in one row, and the PBS tube (117), the primer tube (118), the dye tube (119) and the ddH2O tube (120) are sequentially arranged in another row.

4. A device for virus and bacteria typing detection according to claim 2 or 3, characterized in that: The bottom of the eluent tube (114) is a pointed bottom that protrudes downward.

5. A device for virus and bacteria typing detection according to claim 2 or 3, characterized in that: The tube bodies of the lysate tube (111), the first washing solution tube (112), the second washing solution tube (113), the PBS tube (117), the primer tube (118), the dye tube (119) and the ddH2O tube (120) all have flat bottoms.

6. The device for virus and bacteria typing detection according to claim 1, characterized in that: The microfluidic amplification detection component comprises a microfluidic chip (7), a heating plate (8), a sample addition hole (221), a virus reaction hole and a bacteria reaction hole; the microfluidic chip (7) is provided with a sample addition hole (221), a virus reaction hole and a bacteria reaction hole, the sample addition hole (221) is respectively connected to the virus reaction hole and the bacteria reaction hole, and a heating plate (8) is installed under the microfluidic chip (7).

7. A device for virus and bacteria typing detection according to claim 6, characterized in that: The virus reaction wells include one or more of influenza B virus reaction wells (222), respiratory syncytial virus reaction wells (223), human adenovirus reaction wells (224), influenza A (H1N1) virus reaction wells (225), and influenza A (H7N9) virus reaction wells (226).

8. The device for virus and bacteria typing detection according to claim 6, characterized in that: The bacterial reaction wells include one or more of a Staphylococcus aureus reaction well (227), an Escherichia coli reaction well (228), a Streptococcus pneumoniae reaction well (229), a Klebsiella pneumoniae reaction well (230), and a Pseudomonas aeruginosa reaction well (231).

9. The device for virus and bacteria typing detection according to claim 6, characterized in that: The heating plate (8) is a disposable heating plate and can maintain a constant temperature of 65 degrees Celsius for 30 minutes.

10. The device for virus and bacteria typing detection according to claim 1, characterized in that: The box cover (11) is provided with a groove (10) inside, and the barrel (5) of the syringe can be embedded in the groove (10).

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