Inhalable mouthpiece for detecting target nucleic acids in exhaled breath

Abstract

This invention discloses an embeddable micro-device for detecting target nucleic acids in exhaled air, comprising a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module. The nucleic acid signal amplification module includes a virus attachment strip, one end of which is connected to a solution storage bubble. Six rectangular plates are connected sequentially; each base membrane has a hydrophobic region sealed with wax outside its circle, and within each circle are sequentially arranged nucleic acid lysis buffer, a hydrophilic polymer aqueous solution, an amplification reagent, a hydrophilic polymer aqueous solution, a cleavage reagent, and another hydrophilic polymer aqueous solution. The other end of the virus attachment strip is connected to a first rectangular plate, and a sixth rectangular plate is connected to a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. This invention can be directly used to detect viruses in exhaled droplets. This device achieves effective enrichment of viruses in gas by enriching the gas, enabling highly sensitive, simple, and rapid detection of low concentrations of viruses.

Description

Technical Field

[0001] This invention relates to the field of exhaled breath nucleic acid detection technology, and more specifically to a miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled breath. Background Technology

[0002] Currently, numerous measures have been implemented for the detection of viruses such as SARS-CoV-2 and influenza. Real-time quantitative PCR (qPCR) technology plays a crucial role in this detection, achieving specific signal amplification and improving sensitivity and accuracy by designing primers at highly conserved gene locations. However, it requires highly trained operators, expensive equipment, and certified laboratories, which remains a significant challenge in remote areas lacking resources and medical facilities. Furthermore, the potential for transmission during the time between sampling and obtaining test results for large-scale screening of target viruses cannot be ignored. Therefore, the development of a simple, rapid, environmentally friendly, highly sensitive, and easily embeddable micro-device in a face mask that can directly capture bacteria or viruses from exhaled breath and detect their target nucleic acid sequences based on their gene sequences would provide effective protection during the testing process while rapidly obtaining results. Summary of the Invention

[0003] The purpose of this invention is to overcome the shortcomings of the prior art and provide a miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled air.

[0004] A second objective of this invention is to provide a second micro-device that can be embedded in a mask for detecting target nucleic acids in exhaled breath.

[0005] The technical solution of this invention is summarized as follows:

[0006] An embeddable, mask-style micro-device for detecting target nucleic acids in exhaled breath includes a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module. The sample enrichment module includes an elliptical funnel-shaped cylinder 1, the small end of which is connected to the edge of the elliptical bottom wall 2. A rectangular hole 18 is provided on the bottom wall of the elliptical cylinder. The nucleic acid signal amplification module includes a virus attachment strip 4, which includes a rapidly diffusing material layer 49. A water-absorbing and breathable material layer 47 is provided in the middle of the upper surface of the rapidly diffusing material layer 49. A polymer waterproof and breathable material layer 48 is disposed in the middle of the lower surface of material layer 49, and a solution storage bubble 3 is connected to one end of the virus attachment strip; the solution storage bubble is composed of medical polyvinyl chloride as the bubble body and medical aluminum foil as the substrate, and a storage solution and a needle 50 are disposed inside the solution storage bubble; the first rectangular sheet 51 includes a first base film 5, and a circle with a diameter of 0.35-0.8 cm is drawn in the middle of the first base film, which is the first circle 6. The outside of the first circle is sealed with wax as a hydrophobic area, and 3-40 μL of nucleic acid lysis buffer is disposed inside the first circle. After drying, it is the nucleic acid lysis area; the second rectangular sheet 52 includes a second base film 5. Membrane 7, with a circle of 0.2-0.7 cm in diameter drawn in the middle of the second base membrane, designated as the second circle 8. The area outside the second circle is sealed with wax as a hydrophobic region, and 4-20 μL of hydrophilic polymer aqueous solution is placed inside the second circle. After drying, this forms the first lag region. The third rectangular sheet 53 includes the third base membrane 9, with a circle of 0.35-0.8 cm in diameter drawn in the middle of the third base membrane, designated as the third circle 10. The area outside the third circle is sealed with wax as a hydrophobic region, and 3-40 μL of amplification reagent is placed inside the third circle. After drying, this forms the nucleic acid amplification region. The fourth rectangular sheet 54 includes the fourth base membrane 11, with the fourth base membrane... A circle with a diameter of 0.2-0.7 cm is drawn in the middle, designated as the fourth circle 12. The area outside the fourth circle is sealed with wax as a hydrophobic region. Inside the fourth circle, 4-20 μL of a hydrophilic polymer aqueous solution is placed. After drying, this forms the second lag region. The fifth rectangular sheet 55 includes the fifth base film 13. A circle with a diameter of 0.35-0.8 cm is drawn in the middle of the fifth base film, designated as the fifth circle 14. The area outside the fifth circle is sealed with wax as a hydrophobic region. Inside the fifth circle, 3-40 μL of cutting reagent is placed. After drying, this forms the probe cutting region. The sixth rectangular sheet 56 includes the sixth base film 15. A circle with a diameter of 0.2-0.7 cm is drawn in the middle of the sixth base film.A 7cm circle, designated as the sixth circle 16, has a hydrophobic region sealed with wax outside and contains 4-20μL of a hydrophilic polymer aqueous solution, which, after drying, forms the third lag region. The signal display and detection module is a universal lateral flow test strip 17 for detecting biotin and FITC-labeled analytes. The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular sheet 51. The first rectangular sheet 51, second rectangular sheet 52, third rectangular sheet 53, fourth rectangular sheet 54, fifth rectangular sheet 55, and sixth rectangular sheet 56 may or may not be connected. The third lag region of the sixth rectangular sheet 56 is connected to the universal lateral flow test strip 17 for detecting biotin and FITC-labeled analytes. The nucleic acid signal amplification module and the signal display and detection module cover the outer surface of the elliptical cylindrical bottom wall 2, exposing the polymer waterproof and breathable material layer 48 of the virus attachment strip at the rectangular holes on the elliptical cylindrical bottom wall.

[0007] The second type is an embeddable mask-style micro-device for detecting target nucleic acids in exhaled breath, comprising a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module. The sample enrichment module includes an elliptical funnel-shaped cylinder 101, the small end of which is connected to the edge of the elliptical bottom wall 102. A rectangular hole 118 is provided on the bottom wall of the elliptical cylinder. The nucleic acid signal amplification module includes a virus attachment strip 104, which includes a rapidly diffusing material layer 149. A water-absorbing and breathable material layer 147 is provided in the middle of the upper surface of the rapidly diffusing material layer 149, and a polymer waterproof and breathable material layer 148 is provided in the middle of the lower surface of the rapidly diffusing material layer 149. One end of the virus attachment strip is connected to a solution storage bubble 103; the solution storage bubble is composed of a medical polyvinyl chloride bubble body and a medical aluminum foil substrate, and a storage solution and a needle 150 are disposed inside the solution storage bubble; the first rectangular sheet 151 includes a first base membrane 105, with a circular hole of 0.3-0.75 cm in diameter punched in the middle of the first base membrane, called the first circular hole 106, and the edge of a first circular substrate 140 containing 3-40 μL of nucleic acid lysis buffer with a diameter of 0.35-0.8 cm is attached to the edge of the first circular hole, and after drying, it forms a nucleic acid lysis zone; the second rectangular sheet 152 includes a second base membrane 107, with a circular hole of 0.2-0.7 cm in diameter punched in the middle of the second base membrane, called the second circular hole 108, for storing the virus. A second base membrane with a second circular hole is placed on a hydrophobic material. 4-20 μL of a hydrophilic polymer aqueous solution is dropped into the second circular hole. After drying, a first dense membrane 141 is formed inside and at the edge of the circular hole, constituting a first lag region. A third rectangular sheet 153 includes a third base membrane 109, with a circular hole 110 of 0.3-0.75 cm in diameter punched in the center of the third base membrane. A second circular substrate (142) containing 3-40 μL of amplification reagent, with a diameter of 0.35-0.8 cm, is attached to the edge of the third circular hole. After drying, this forms a nucleic acid amplification region. A fourth rectangular sheet 154 includes a fourth base membrane 111, with a circular hole 110 of 0.2-0.7 cm in diameter punched in the center of the fourth base membrane. The fourth circular hole 112 is formed by placing the fourth base film with the fourth circular hole on a hydrophobic material and adding 4-20 μL of a hydrophilic polymer aqueous solution into the fourth circular hole. After drying, a second dense film 143 is formed inside and at the edge of the circular hole, which is the second lag region. The fifth rectangular sheet 155 includes the fifth base film 113, and a circular hole with a diameter of 0.3-0.75 cm is punched in the middle of the fifth base film, which is the fifth circular hole 114. The edge of the third circular substrate 144 with a diameter of 0.35-0.8 cm containing 3-40 μL of cutting reagent is attached to the edge of the fifth circular hole. After drying, it is the probe cutting region. The sixth rectangular sheet 156 includes the sixth base film 115, and a hole with a diameter of 0.2-0.8 cm is punched in the middle of the sixth base film.A 7cm circular hole, designated as the sixth circular hole 116, is placed on a hydrophobic material. 4-20μL of a hydrophilic polymer aqueous solution is added to the sixth circular hole. After drying, a third dense film 145, representing the third lag region, is formed inside and around the circular hole. The signal display and detection module is a universal lateral flow test strip 117 used for detecting biotin and FITC-labeled analytes. The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular sheet 151. The second rectangular strip 152, the third rectangular strip 153, the fourth rectangular strip 154, the fifth rectangular strip 155, and the sixth rectangular strip 156 may be interconnected or not connected; the third lag zone of the sixth rectangular strip 156 is connected to a universal lateral flow test strip 117 used for detecting biotin and FITC-labeled analytes; a nucleic acid signal amplification module and a signal display detection module cover the outer surface of the bottom wall 102 of the elliptical cylinder; and the polymer waterproof and breathable material layer 148 of the virus attachment strip is exposed at the rectangular holes on the bottom wall of the elliptical cylinder.

[0008] Preferably, the storage solution consists of 1-5 T / mL of ribonuclease inhibitor, 0.01%-0.05% TritiumX-100, 0.5%-2.5% CHAPS, and 0.01%-0.05% NP-40, with the remainder being nuclease-free water.

[0009] Preferably, the material of the rapid diffusion material layer is cotton cloth or filter paper, and the thickness of the rapid diffusion material layer is 0.02 to 0.5 mm.

[0010] Preferably, the absorbent and breathable material layer is made of cellulose paper, nylon fiber cloth or polyester fiber cloth; the thickness of the absorbent and breathable material layer is 0.01 to 0.5 mm.

[0011] Preferably, the polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the polymer waterproof and breathable material layer is 0.01 to 0.5 mm.

[0012] Preferably, the nucleic acid lysis buffer consists of: 0.001%–0.05% NP-40, 1%–3% CHAPS, 0.5%–3% lysozyme, 2%–10% sucrose, and the remainder being 10–100 mM Tris buffer at pH 7.5.

[0013] Preferably, the aqueous solution of the hydrophilic polymer has a concentration of 4wt%-20wt%; the hydrophilic polymer is polyvinyl alcohol with a weight average molecular weight of 30,000-100,000, polyethylene glycol with a weight average molecular weight of 50,000-150,000, polypropylene glycol with a weight average molecular weight of 50,000-150,000, sodium carboxymethyl cellulose with a weight average molecular weight of 30,000-250,000, or sodium alginate with a weight average molecular weight of 10,000-20,000.

[0014] Preferably, the amplification reagents are: TvsX 50–300 ng / μL, TvsY 30–200 ng / μL, T4 gp32 500–2000 ng / μL, polymerase 20–150 ng / μL, creatine kinase 50–450 ng / μL, RNase inhibitor 0.02–50 ng / μL, AMV reverse transcriptase 0.02–50 ng / μL, RNase H 0.2–80 ng / μL, Tris buffer 20–100 mM, potassium acetate 70–120 mM, magnesium acetate 10–20 mM, dithiothreitol 1–4 mM, PEG 4 wt%–10 wt%, dNTP 300–800 μM, and ATP. 2–6 mM, creatine phosphate 30–80 mM, BSA 0.01 wt%–5 wt%, primers 0.1–4 μM, the polymerase being Bst polymerase or Sat polymerase; the PEG being PEG 20000 or PEG 35000.

[0015] Preferably, the cleavage reagent is: 3-20 mM magnesium salt, 0.01 wt%-5% BSA, 40-2000 ng / μL Cas12a, 1-10 mM DTT, 0.2-1 pmol FB probe, and 0.05-3 μM viral gene targeting gRNA; the balance is 3-50 mM Tris-HCl buffer at pH 7.0-8.5; wherein the FB probe is a single-stranded nucleic acid sequence labeled with FITC fluorescent pigment at the 5' end and biotin at the 3' end, and the magnesium salt is magnesium chloride or magnesium acetate.

[0016] Preferably, the thickness of the first base membrane 5, the second base membrane 7, the third base membrane 9, the fourth base membrane 11, the fifth base membrane 13, and the sixth base membrane 15 of the first embeddable mask-type micro-device for detecting target nucleic acids in exhaled gas is 0.01 to 0.5 mm, and the material is cellulose paper, cotton cloth, nylon fiber cloth or polyester fiber cloth, and the wax is microcrystalline wax, paraffin wax, petrolatum or polypropylene.

[0017] Preferably, the thickness of the first base membrane 105, the second base membrane 107, the third base membrane 109, the fourth base membrane 111, the fifth base membrane 113, and the sixth base membrane 115 of the second type of embeddable mask-type micro-device for detecting target nucleic acids in exhaled gas is 20-90 μm, and the material is biaxially oriented polypropylene transparent tape, butyl waterproof self-adhesive aluminum foil tape, or resin pressure-sensitive double-sided tape.

[0018] Advantages of this invention:

[0019] This invention can be directly used to detect viruses in exhaled droplets. The device effectively enriches exhaled gases to detect viruses, enabling highly sensitive, simple, and rapid detection of low concentrations of viruses in exhaled gases. Attached Figure Description

[0020] Figure 1 This is a schematic diagram of the first embeddable micro-device for detecting target nucleic acids in exhaled breath;

[0021] Figure 2 This is a schematic diagram of a sample enrichment module embedded in a mask-style micro-device for detecting target nucleic acids in exhaled breath;

[0022] Figure 3 This is a top view schematic diagram of the nucleic acid signal amplification module and signal display detection module in the first type of micro-device that can be embedded in a mask for detecting target nucleic acids in exhaled air;

[0023] Figure 4 This is a schematic diagram showing the interconnection of the storage bubble, virus attachment strip, first rectangular plate, second rectangular plate, third rectangular plate, fourth rectangular plate, fifth rectangular plate, and sixth rectangular plate in the nucleic acid signal amplification module of the first embeddable micro-device for detecting target nucleic acids in exhaled gas.

[0024] Figure 5 This is a schematic diagram of the composition of the first virus attachment strip and the solution storage bubble used to detect target nucleic acids in exhaled breath;

[0025] Figure 6 This is a schematic diagram of a second type of micro-device that can be embedded in a mask for detecting target nucleic acids in exhaled breath;

[0026] Figure 7 This is a schematic diagram of a sample enrichment module that can be embedded in a mask-style micro-device for detecting target nucleic acids in exhaled breath;

[0027] Figure 8 This is a top view schematic diagram of the nucleic acid signal amplification module and signal display detection module in a second type of micro-device that can be embedded in a mask for detecting target nucleic acids in exhaled air;

[0028] Figure 9 This is a schematic diagram showing the interconnection of the storage bubble, virus attachment strip, first rectangular plate, second rectangular plate, third rectangular plate, fourth rectangular plate, fifth rectangular plate, and sixth rectangular plate in the nucleic acid signal amplification module of the second type of embeddable mask-type micro-device for detecting target nucleic acids in exhaled gas;

[0029] Figure 10 This is a schematic diagram of the composition of the virus attachment strip and the solution storage bubble for the second type of virus attachment strip used to detect target nucleic acids in exhaled breath;

[0030] Figure 11 To detect the virus using the first embeddable microdevice for detecting target nucleic acids in exhaled breath, wherein... Figure 11A represents the result of the COVID-19 test; Figure 11 B represents the result of the detection of influenza A virus; Figure 11 C represents the result of the detection of influenza B virus;

[0031] Figure 12 To detect the virus using a second type of embeddable micro-device for detecting target nucleic acids in exhaled breath, wherein... Figure 12 A represents the result of the COVID-19 test; Figure 12 B represents the result of the detection of influenza A virus; Figure 12 C represents the result of detecting influenza B virus. Detailed Implementation

[0032] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments, but the scope of protection of the present invention is not limited thereto. Equivalent substitutions or corresponding improvements made to the content of the present invention shall still fall within the scope of protection of the present invention.

[0033] Example 1

[0034] The storage solution consists of 3 T / mL of ribonuclease inhibitor (human placental gene source), 0.03% TritiumX-100, 1.5% CHAPS, and 0.03% NP-40, with the remainder being nuclease-free water.

[0035] Example 2

[0036] The storage solution consists of 1 T / mL of ribonuclease inhibitor (human placental gene source), 0.05% TritiumX-100, 0.5% CHAPS, and 0.05% NP-40, with the remainder being nuclease-free water.

[0037] Example 3

[0038] The storage solution consisted of 5 T / mL of ribonuclease inhibitor (derived from rat liver genes), 0.01% TritiumX-100, 2.5% CHAPS, and 0.01% NP-40, with the remainder being nuclease-free water.

[0039] Example 4: The composition of the nucleic acid lysis buffer is as follows:

[0040] The composition consists of 0.01% NP-40, 1% CHAPS, 1% lysozyme, 5% sucrose, and the remainder is 50 mM Tris buffer at pH 7.5.

[0041] Example 5: The composition of the nucleic acid lysis buffer is as follows:

[0042] The composition consists of 0.001% NP-40, 3% CHAPS, 0.5% lysozyme, 10% sucrose, and the balance being 10 mM Tris buffer at pH 7.5.

[0043] Example 6: The composition of the nucleic acid lysis buffer is as follows:

[0044] The composition consists of 0.05% NP-40, 2% CHAPS, 3% lysozyme, 2% sucrose, and the remainder is 100 mM Tris buffer at pH 7.5.

[0045] Example 7

[0046] Aqueous solution of hydrophilic polymer: polyvinyl alcohol with a concentration of 12 wt% and a weight average molecular weight of 80,000.

[0047] Experiments have shown that replacing the polyvinyl alcohol in this embodiment with polyvinyl alcohol at a concentration of 4 wt% and a weight-average molecular weight of 100,000 or with polyvinyl alcohol at a concentration of 20 wt% and a weight-average molecular weight of 30,000 yields the corresponding hydrophilic polymer aqueous solutions.

[0048] Example 8

[0049] Aqueous solution of hydrophilic polymer: 4 wt% polypropylene glycol with a weight average molecular weight of 100,000.

[0050] Experiments have shown that replacing the polypropylene glycol in this embodiment with polypropylene glycol at a concentration of 10 wt% (weight average molecular weight 150,000), or polypropylene glycol at a concentration of 20 wt% (weight average molecular weight 50,000), or polyethylene glycol at a concentration of 10 wt% (weight average molecular weight 150,000), or polyethylene glycol at a concentration of 20 wt% (weight average molecular weight 50,000), or polyethylene glycol at a concentration of 4 wt% (weight average molecular weight 100,000), yields the corresponding hydrophilic polymer aqueous solutions.

[0051] Example 9

[0052] Aqueous solution of hydrophilic polymer: 20wt% sodium carboxymethyl cellulose with a weight average molecular weight of 100,000.

[0053] Experiments have shown that replacing the sodium carboxymethyl cellulose in this embodiment with 4 wt% sodium carboxymethyl cellulose with a weight average molecular weight of 250,000, 20 wt% sodium carboxymethyl cellulose with a weight average molecular weight of 30,000, 4 wt% sodium alginate with a weight average molecular weight of 10,000, 20 wt% sodium alginate with a weight average molecular weight of 10,000, and 10 wt% sodium alginate with a weight average molecular weight of 20,000 yields the corresponding hydrophilic polymer aqueous solution.

[0054] Example 10

[0055] Amplification reagent composition: TvsX 200ng / μL, TvsY 120ng / μL, T4 gp32 1200ng / μL, Bst polymerase 90ng / μL, creatine kinase 300ng / μL, RNase inhibitor (human placental gene source) 25ng / μL, AMV reverse transcriptase 25ng / μL, RNase H 40ng / μL, Tris buffer 50mM, potassium acetate 100mM, magnesium acetate 15mM, dithiothreitol 2mM, PEG30000 6wt%, dNTP 600μM, ATP 4mM, creatine phosphate 60mM, BSA 3wt%, primers (primers for the SARS-CoV2 S protein gene, in a molar ratio of 1:1:1, SEQ ID No. 2, SEQ ID No. 3, SEQ ID No. 4) 5μM.

[0056] Example 11

[0057] Amplification reagent composition: TvsX 50 ng / μL, TvsY 200 ng / μL, T4 gp32 500 ng / μL, Sat polymerase 150 ng / μL, creatine kinase 50 ng / μL, RNase inhibitor (human placental gene source) 50 ng / μL, AMV reverse transcriptase 0.02 ng / μL, RNase H 80 ng / μL, Tris buffer 20 mM, potassium acetate 120 mM, magnesium acetate 10 mM, dithiothreitol 4 mM, PEG 35000 4 wt%, dNTP 800 μM, ATP 2 mM, creatine phosphate 80 mM, BSA 0.01 wt%, primers (primers for the matrix protein 2 gene of influenza A virus, in a molar ratio of 1:1:1, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10) 10 μM.

[0058] Example 12

[0059] Composition of amplification reagents:

[0060] TvsX 300 ng / μL, TvsY 30 ng / μL, T4 gp32 2000 ng / μL, Bst polymerase 20 ng / μL, creatine kinase 450 ng / μL, RNase inhibitor (rat liver gene source) 0.02 ng / μL, AMV reverse transcriptase 50 ng / μL, RNase H 0.2 ng / μL, Tris buffer 100 mM, potassium acetate 70 mM, magnesium acetate 20 mM, dithiothreitol 1 mM, PEG 20000 10 wt%, dNTP 300 μM, ATP 6 mM, creatine phosphate 30 mM, BSA 5 wt%, primers (primers for the matrix protein 2 gene of influenza B virus: SEQ ID No. 14, SEQ ID No. 15, SEQ ID No. 16 in a molar ratio of 1:1:1) 1 μM.

[0061] Example 13

[0062] Composition of the cutting reagent:

[0063] Magnesium chloride 10mM, BSA 3%, Cas12a 1000ng / μL, DTT 5mM, FB probe 0.5pmol, viral gene targeting gRNA (SEQ ID No.7) 2μM; the balance is 25mM Tris-HCl buffer at pH 8.

[0064] The FB probe is a single-stranded nucleic acid sequence labeled with FITC at the 5' end and biotin at the 3' end: (5'-6FAM / ATTATTTTATTA / 3'Biotin).

[0065] Example 14

[0066] Composition of the cutting reagent:

[0067] Magnesium chloride 3mM, BSA 5%, Cas12a 40ng / μL, DTT 10mM, FB probe 0.2pmol, viral gene targeting gRNA 3μM (SEQ ID No. 13); the balance was 50mM Tris-HCl buffer at pH 7.0.

[0068] The FB probe is a single-stranded nucleic acid sequence labeled with FITC at the 5' end and biotin at the 3' end: (5'-6FAM / ATTATTTTATTA / 3'Biotin).

[0069] Example 15

[0070] Composition of the cutting reagent:

[0071] Magnesium acetate 20mM, BSA 0.01wt%, Cas12a 2000ng / μL, DTT 1mM, FB probe 1pmol, viral gene targeting gRNA 0.05μM (SEQ ID No.1); the balance is 3mM Tris-HCl buffer at pH 8.5.

[0072] The FB probe is a single-stranded nucleic acid sequence labeled with FITC at the 5' end and biotin at the 3' end (5'-6FAM / ATTATTTTATTA / 3'Biotin).

[0073] Example 16

[0074] The first type of miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled breath, see [link / description]. Figure 1 and Figure 2 It includes a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module;

[0075] The sample enrichment module includes an elliptical funnel-shaped cylinder 1, the small end of which is connected to the edge of the bottom wall 2 of the elliptical cylinder, and a rectangular hole 18 is provided on the bottom wall of the elliptical cylinder.

[0076] The nucleic acid signal amplification module (see Figure 3 , Figure 4 , Figure 5 The virus attachment strip 4 includes a rapid diffusion material layer 49, a water-absorbing and breathable material layer 47 is provided in the middle of the upper surface of the rapid diffusion material layer 49, a polymer waterproof and breathable material layer 48 is provided in the middle of the lower surface of the rapid diffusion material layer 49, and a solution storage bubble 3 is connected to one end of the virus attachment strip.

[0077] The solution storage bubble is composed of a medical-grade polyvinyl chloride bubble body connected to a medical-grade aluminum foil substrate. The storage solution of Example 1 and the needle 50 are placed inside the solution storage bubble.

[0078] The first rectangular sheet 51 includes a first base film 5, and a circle with a diameter of 0.6 cm is drawn in the middle of the first base film, which is the first circle 6. The outside of the first circle is sealed with wax as a hydrophobic area, and 30 μL of the nucleic acid lysis buffer of Example 4 is placed inside the first circle. After drying, it is the nucleic acid lysis area.

[0079] The second rectangular sheet 52 includes a second base film 7, and a circle with a diameter of 0.4 cm is drawn in the middle of the second base film, which is the second circle 8. The area outside the second circle is sealed with wax as a hydrophobic region, and 10 μL of the hydrophilic polymer aqueous solution of Example 7 is placed inside the second circle. After drying, it is the first lag region.

[0080] The third rectangular sheet 53 includes a third base membrane 9, and a circle with a diameter of 0.6 cm is drawn in the middle of the third base membrane, which is the third circle 10. The area outside the third circle is sealed with a hydrophobic region, and 30 μL of the amplification reagent of Example 10 is placed inside the third circle. After drying, it is the nucleic acid amplification region.

[0081] The fourth rectangular sheet 54 includes a fourth base film 11, with a 0.4 cm circle drawn in the middle of the fourth base film, which is the fourth circle 12. The area outside the fourth circle is sealed with wax as a hydrophobic region, and 10 μL of the hydrophilic polymer aqueous solution of Example 7 is placed inside the fourth circle. After drying, it is the second lag region.

[0082] The fifth rectangular sheet 55 includes a fifth base film 13. A circle with a diameter of 0.6 cm is drawn in the middle of the fifth base film, which is the fifth circle 14. The area outside the fifth circle is sealed with wax as a hydrophobic area, and 30 μL of the cutting reagent of Example 13 is placed inside the fifth circle. After drying, it becomes the probe cutting area.

[0083] The sixth rectangular sheet 56 includes a sixth base film 15, with a 0.4 cm circle drawn in the middle of the sixth base film, which is the sixth circle 16. The area outside the sixth circle is sealed with wax as a hydrophobic region, and 10 μL of the hydrophilic polymer aqueous solution of Example 7 is placed inside the sixth circle. After drying, it is the third lag region.

[0084] The signal display detection module is a commercially available lateral flow test strip 17 for detecting biotin and FITC-labeled analytes;

[0085] The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular piece 51, and the first rectangular piece 51, the second rectangular piece 52, the third rectangular piece 53, the fourth rectangular piece 54, the fifth rectangular piece 55 and the sixth rectangular piece 56 are interconnected;

[0086] The third lag zone of the sixth rectangular strip 56 is connected to the universal lateral flow test strip 17 for detecting biotin and FITC-labeled analytes;

[0087] The nucleic acid signal amplification module and the signal display and detection module cover the outer surface of the bottom wall 2 of the elliptical cylinder; and expose the polymer waterproof and breathable material layer 48 of the virus attachment strip at the rectangular hole on the bottom wall of the elliptical cylinder.

[0088] The rapid diffusion material layer is made of cotton cloth and has a thickness of 0.3 mm.

[0089] The absorbent and breathable material layer is made of cellulose paper; the thickness of the absorbent and breathable material layer is 0.3mm.

[0090] The polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the polymer waterproof and breathable material layer is 0.3mm.

[0091] The first type of embeddable mask-type micro-device for detecting target nucleic acids in exhaled air has a thickness of 0.3 mm for its first base membrane 5, second base membrane 7, third base membrane 9, fourth base membrane 11, fifth base membrane 13, and sixth base membrane 15, and is made of cellulose paper.

[0092] The wax is a microcrystalline wax.

[0093] Example 17

[0094] The first type of miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled breath, see [link / description]. Figure 1 and Figure 2 It includes a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module; the sample enrichment module includes an elliptical funnel-shaped cylinder 1, the small end of which is connected to the edge of the bottom wall 2 of the elliptical cylinder, and a rectangular hole 18 is provided on the bottom wall of the elliptical cylinder; the nucleic acid signal amplification module (see...) Figure 3 , Figure 4 , Figure 5 The device includes a virus attachment strip 4, which includes a rapid diffusion material layer 49. A water-absorbing and breathable material layer 47 is disposed in the middle of the upper surface of the rapid diffusion material layer 49, and a polymer waterproof and breathable material layer 48 is disposed in the middle of the lower surface of the rapid diffusion material layer 49. A solution storage bubble 3 is connected to one end of the virus attachment strip. The solution storage bubble is composed of medical polyvinyl chloride as the bubble body and medical aluminum foil as the substrate. The storage solution of Example 2 and the needle 50 are disposed inside the solution storage bubble. The first rectangular sheet 51 includes a first base film 5. A circle with a diameter of 0.35 cm is drawn in the middle of the first base film, which is the first circle 6. The outside of the first circle is sealed with wax as a hydrophobic area. 3 μL of nucleic acid lysis buffer of Example 5 is disposed inside the first circle. After drying, it is the nucleic acid lysis area.

[0095] The second rectangular sheet 52 includes a second base film 7, and a circle with a diameter of 0.2 cm is drawn in the middle of the second base film, which is the second circle 8. The area outside the second circle is sealed with wax as a hydrophobic region, and 4 μL of the hydrophilic polymer aqueous solution of Example 8 is placed inside the second circle. After drying, it is the first lag region.

[0096] The third rectangular sheet 53 includes a third base membrane 9, and a circle with a diameter of 0.35 cm is drawn in the middle of the third base membrane, which is the third circle 10. The area outside the third circle is sealed with a hydrophobic region, and 3 μL of the amplification reagent of Example 11 is placed inside the third circle. After drying, it is the nucleic acid amplification region.

[0097] The fourth rectangular sheet 54 includes a fourth base film 11, with a 0.2 cm circle drawn in the middle of the fourth base film, which is the fourth circle 12. The area outside the fourth circle is sealed with wax as a hydrophobic region, and 4 μL of the hydrophilic polymer aqueous solution of Example 8 is placed inside the fourth circle. After drying, it is the second lag region.

[0098] The fifth rectangular sheet 55 includes a fifth base film 13. A circle with a diameter of 0.35 cm is drawn in the middle of the fifth base film, which is the fifth circle 14. The area outside the fifth circle is sealed with wax as a hydrophobic area, and 3 μL of the cutting reagent of Example 14 is placed inside the fifth circle. After drying, it becomes the probe cutting area.

[0099] The sixth rectangular sheet 56 includes a sixth base film 15, with a 0.2 cm circle drawn in the middle of the sixth base film, which is the sixth circle 16. The area outside the sixth circle is sealed with wax as a hydrophobic region, and 4 μL of the hydrophilic polymer aqueous solution of Example 8 is placed inside the sixth circle. After drying, it is the third lag region.

[0100] The signal display detection module is a commercially available lateral flow test strip 17 for detecting biotin and FITC-labeled analytes;

[0101] The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular piece 51. The first rectangular piece 51, the second rectangular piece 52, the third rectangular piece 53, the fourth rectangular piece 54, the fifth rectangular piece 55 and the sixth rectangular piece 56 are not connected to each other.

[0102] The third lag zone of the sixth rectangular strip 56 is connected to the universal lateral flow test strip 17 for detecting biotin and FITC-labeled analytes;

[0103] The nucleic acid signal amplification module and the signal display and detection module cover the outer surface of the bottom wall 2 of the elliptical cylinder; and expose the polymer waterproof and breathable material layer 48 of the virus attachment strip at the rectangular hole on the bottom wall of the elliptical cylinder.

[0104] The rapid diffusion material layer is made of cotton cloth and has a thickness of 0.02 mm.

[0105] The absorbent and breathable material layer is made of nylon fiber cloth; the thickness of the absorbent and breathable material layer is 0.01mm.

[0106] The polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the polymer waterproof and breathable material layer is 0.01mm.

[0107] The thickness of the first base film 5, the second base film 7, the third base film 9, the fourth base film 11, the fifth base film 13, and the sixth base film 15 is 0.01 mm, and the material is nylon fiber cloth.

[0108] The wax is paraffin wax.

[0109] Example 18

[0110] The first type of miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled breath, see [link / description]. Figure 1 and Figure 2It includes a sample enrichment module, characterized in that it also includes a nucleic acid signal amplification module and a signal display and detection module;

[0111] The sample enrichment module includes an elliptical funnel-shaped cylinder 1, the small end of which is connected to the edge of the bottom wall 2 of the elliptical cylinder, and a rectangular hole 18 is provided on the bottom wall of the elliptical cylinder.

[0112] The nucleic acid signal amplification module (see Figure 3 , Figure 4 , Figure 5 )include

[0113] The virus attachment strip 4 and the pathogen attachment strip include a fast-diffusion material layer 49, a water-absorbing and breathable material layer 47 is provided in the middle of the upper surface of the fast-diffusion material layer 49, a polymer waterproof and breathable material layer 48 is provided in the middle of the lower surface of the fast-diffusion material layer 49, and a solution storage bubble 3 is connected to one end of the virus attachment strip.

[0114] The solution storage bubble is composed of a medical polyvinyl chloride bubble body and a medical aluminum foil substrate. The storage solution of Example 3 and the needle 50 are disposed inside the solution storage bubble.

[0115] The first rectangular sheet 51 includes a first base film 5, and a circle with a diameter of 0.8 cm is drawn in the middle of the first base film, which is the first circle 6. The area outside the first circle is sealed with wax as a hydrophobic area, and 40 μL of the nucleic acid lysis buffer from Example 6 is placed inside the first circle. After drying, it is the nucleic acid lysis area.

[0116] The second rectangular sheet 52 includes a second base film 7, and a circle with a diameter of 0.7 cm is drawn in the middle of the second base film, which is the second circle 8. The area outside the second circle is sealed with wax as a hydrophobic region, and 20 μL of the hydrophilic polymer aqueous solution of Example 9 is placed inside the second circle. After drying, it is the first lag region.

[0117] The third rectangular sheet 53 includes a third base membrane 9, and a circle with a diameter of 0.8 cm is drawn in the middle of the third base membrane, which is the third circle 10. The area outside the third circle is sealed with a hydrophobic region, and 40 μL of the amplification reagent of Example 12 is placed inside the third circle. After drying, it is the nucleic acid amplification region.

[0118] The fourth rectangular sheet 54 includes a fourth base film 11, with a 0.7 cm circle drawn in the middle of the fourth base film, which is the fourth circle 12. The area outside the fourth circle is sealed with wax as a hydrophobic region, and 20 μL of the hydrophilic polymer aqueous solution of Example 9 is placed inside the fourth circle. After drying, it is the second lag region.

[0119] The fifth rectangular sheet 55 includes a fifth base film 13. A circle with a diameter of 0.8 cm is drawn in the middle of the fifth base film, which is the fifth circle 14. The area outside the fifth circle is sealed with wax as a hydrophobic area. 40 μL of the cutting reagent of Example 15 is placed inside the fifth circle. After drying, it is the probe cutting area.

[0120] The sixth rectangular sheet 56 includes a sixth base film 15, with a 0.7 cm circle drawn in the middle of the sixth base film, which is the sixth circle 16. The area outside the sixth circle is sealed with wax as a hydrophobic region, and 20 μL of the hydrophilic polymer aqueous solution of Example 9 is placed inside the sixth circle. After drying, it is the third lag region.

[0121] The signal display detection module is a commercially available lateral flow test strip 17 for detecting biotin and FITC-labeled analytes;

[0122] The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular piece 51, and the first rectangular piece 51, the second rectangular piece 52, the third rectangular piece 53, the fourth rectangular piece 54, the fifth rectangular piece 55 and the sixth rectangular piece 56 are interconnected;

[0123] The third lag zone of the sixth rectangular strip 56 is connected to the universal lateral flow test strip 17 for detecting biotin and FITC-labeled analytes;

[0124] The nucleic acid signal amplification module and the signal display and detection module cover the outer surface of the bottom wall 2 of the elliptical cylinder; and expose the polymer waterproof and breathable material layer 48 of the virus attachment strip at the rectangular hole on the bottom wall of the elliptical cylinder.

[0125] The rapid diffusion material layer is made of filter paper and has a thickness of 0.5 mm.

[0126] The absorbent and breathable material layer is made of polyester fiber cloth; the thickness of the absorbent and breathable material layer is 0.5mm.

[0127] The polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the polymer waterproof and breathable material layer is 0.5mm.

[0128] The thickness of the first base film 5, the second base film 7, the third base film 9, the fourth base film 11, the fifth base film 13, and the sixth base film 15 is 0.5 mm, and the material is polyester fiber cloth.

[0129] The wax is petrolatum (polypropylene can also be used).

[0130] Example 19

[0131] The second type is a miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled breath. Figure 6 and Figure 7It includes a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module;

[0132] The sample enrichment module includes an elliptical funnel-shaped cylinder 101, the small end of which is connected to the edge of the bottom wall 102 of the elliptical cylinder, and a rectangular hole 118 is provided on the bottom wall of the elliptical cylinder.

[0133] The nucleic acid signal amplification module (see Figure 8 , Figure 9 and Figure 10 The virus attachment strip includes a virus attachment strip 104, which includes a fast-diffusion material layer 149, a water-absorbing and breathable material layer 147 disposed in the middle of the upper surface of the fast-diffusion material layer 149, a polymer waterproof and breathable material layer 148 disposed in the middle of the lower surface of the fast-diffusion material layer 149, and a solution storage bubble 103 connected to one end of the virus attachment strip.

[0134] The solution storage bubble is composed of a medical polyvinyl chloride bubble body connected to a medical aluminum foil substrate. The storage solution of Example 1 and the needle 150 are placed inside the solution storage bubble.

[0135] The first rectangular sheet 151 includes a first base film 105, and a circular hole with a diameter of 0.55 cm is punched in the middle of the first base film, which is the first circular hole 106. The edge of the first circular substrate 140 containing 30 μL of nucleic acid lysis buffer from Example 4, with a diameter of 0.6 cm, is attached to the edge of the first circular hole and dried to form a nucleic acid lysis zone.

[0136] The second rectangular sheet 152 includes a second base film 107, with a circular hole of 0.4 cm in diameter punched in the middle of the second base film, which is called the second circular hole 108. The second base film with the second circular hole is placed on a hydrophobic material (polyethylene plastic film, the same below), and 10 μL of the hydrophilic polymer aqueous solution of Example 7 is dropped into the second circular hole. After drying, a first dense film 141 is formed in the circular hole and at the edge of the circular hole, which is the first lag region.

[0137] The third rectangular sheet 153 includes a third base membrane 109, with a 0.55 cm diameter circular hole punched in the middle of the third base membrane, which is called the third circular hole 110. The edge of the second circular substrate 142 containing 30 μL of the amplification reagent of Example 10, with a diameter of 0.6 cm, is attached to the edge of the third circular hole. After drying, it is a nucleic acid amplification area.

[0138] The fourth rectangular sheet 154 includes a fourth base film 111, with a circular hole of 0.4 cm in diameter punched in the middle of the fourth base film, which is the fourth circular hole 112. The fourth base film with the fourth circular hole is placed on a hydrophobic material, and 10 μL of the hydrophilic polymer aqueous solution of Example 7 is dropped into the fourth circular hole. After drying, a second dense film 143 is formed in the circular hole and at the edge of the circular hole, which is the second lag region.

[0139] The fifth rectangular sheet 155 includes a fifth base film 113, with a 0.55 cm diameter circular hole punched in the middle of the fifth base film, which is the fifth circular hole 114. The edge of the third circular substrate 144 containing 30 μL of the cutting reagent of Example 13 with a diameter of 0.6 cm is attached to the edge of the fifth circular hole and dried to form the probe cutting area.

[0140] The sixth rectangular sheet 156 includes a sixth base film 115, with a circular hole of 0.4 cm in diameter punched in the middle of the sixth base film, which is the sixth circular hole 116. The sixth base film with the sixth circular hole is placed on a hydrophobic material, and 10 μL of the hydrophilic polymer aqueous solution of Example 7 is dropped into the sixth circular hole. After drying, a third dense film 145 is formed inside the circular hole and at the edge of the circular hole, which is the third lag region.

[0141] The signal display detection module is a universal lateral flow test strip 117 for detecting biotin and FITC-labeled analytes;

[0142] The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular piece 151, and the first rectangular piece 151, the second rectangular piece 152, the third rectangular piece 153, the fourth rectangular piece 154, the fifth rectangular piece 155 and the sixth rectangular piece 156 are interconnected.

[0143] The third lag zone of the sixth rectangular strip 156 is connected to a universal lateral flow test strip 117 for detecting biotin and FITC-labeled analytes;

[0144] The nucleic acid signal amplification module and the signal display and detection module cover the outer surface of the bottom wall 102 of the elliptical cylinder; and expose the polymer waterproof and breathable material layer 148 of the virus attachment strip at the rectangular hole on the bottom wall of the elliptical cylinder.

[0145] The rapid diffusion material layer is made of cotton cloth, and the thickness of the rapid diffusion material layer is 0.3 mm.

[0146] The absorbent and breathable material layer is made of cellulose paper; the thickness of the absorbent and breathable material layer is 0.3 mm.

[0147] The polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the waterproof and breathable material layer is 0.3mm.

[0148] The second type of embeddable mask-style micro-device for detecting target nucleic acids in exhaled air has a first base membrane 105, a second base membrane 107, a third base membrane 109, a fourth base membrane 111, a fifth base membrane 113, and a sixth base membrane 115 with a thickness of 70 μm and is made of biaxially oriented polypropylene transparent tape.

[0149] Example 20

[0150] The second type is a miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled breath. Figure 6 and Figure 7 It includes a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module;

[0151] The sample enrichment module includes an elliptical funnel-shaped cylinder 101, the small end of which is connected to the edge of the bottom wall 102 of the elliptical cylinder, and a rectangular hole 118 is provided on the bottom wall of the elliptical cylinder.

[0152] The nucleic acid signal amplification module (see Figure 8 , Figure 9 and Figure 10 )include

[0153] Virus attachment strip 104, the virus attachment strip includes a fast-diffusion material layer 149, a water-absorbing and breathable material layer 147 is provided in the middle of the upper surface of the fast-diffusion material layer 149, a polymer waterproof and breathable material layer 148 is provided in the middle of the lower surface of the fast-diffusion material layer 149, and a solution storage bubble 103 is connected to one end of the virus attachment strip.

[0154] The solution storage bubble is composed of a medical polyvinyl chloride bubble body and a medical aluminum foil substrate. The storage solution of Example 2 and the needle 150 are placed inside the solution storage bubble.

[0155] The first rectangular sheet 151 includes a first base film 105, and a circular hole with a diameter of 0.3 cm is punched in the middle of the first base film, which is the first circular hole 106. The edge of the first circular substrate 140 containing 3 μL of nucleic acid lysis buffer from Example 5, with a diameter of 0.35 cm, is attached to the edge of the first circular hole and dried to form a nucleic acid lysis zone.

[0156] The second rectangular sheet 152 includes a second base film 107, with a circular hole of 0.2 cm in diameter punched in the middle of the second base film, which is called the second circular hole 108. The second base film with the second circular hole is placed on a hydrophobic material, and 4 μL of the hydrophilic polymer aqueous solution of Example 8 is dropped into the second circular hole. After drying, a first dense film 141 is formed in the circular hole and at the edge of the circular hole, which is the first lag region.

[0157] The third rectangular sheet 153 includes a third base membrane 109, with a 0.3 cm diameter circular hole punched in the middle of the third base membrane, which is called the third circular hole 110. The edge of the second circular substrate 142 containing 3 μL of the amplification reagent of Example 11 is attached to the edge of the third circular hole with a diameter of 0.35 cm. After drying, it is a nucleic acid amplification area.

[0158] The fourth rectangular sheet 154 includes a fourth base film 111, with a 0.2 cm diameter circular hole punched in the middle of the fourth base film, which is the fourth circular hole 112. The fourth base film with the fourth circular hole is placed on a hydrophobic material, and 4 μL of the hydrophilic polymer aqueous solution of Example 8 is dropped into the fourth circular hole. After drying, a second dense film 143 is formed inside the circular hole and at the edge of the circular hole, which is the second lag region.

[0159] The fifth rectangular sheet 155 includes a fifth base film 113, with a 0.3 cm diameter circular hole punched in the middle of the fifth base film, which is the fifth circular hole 114. The edge of the third circular substrate 144 containing 3 μL of the cutting reagent of Example 14 with a diameter of 0.35 cm is attached to the edge of the fifth circular hole and dried to form the probe cutting area.

[0160] The sixth rectangular sheet 156 includes a sixth base film 115, with a circular hole of 0.2 cm in diameter punched in the middle of the sixth base film, which is the sixth circular hole 116. The sixth base film with the sixth circular hole is placed on a hydrophobic material, and 4 μL of the hydrophilic polymer aqueous solution of Example 8 is dropped into the sixth circular hole. After drying, a third dense film 145 is formed inside the circular hole and at the edge of the circular hole, which is the third lag region.

[0161] The signal display detection module is a universal lateral flow test strip 117 for detecting biotin and FITC-labeled analytes;

[0162] The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular piece 151. The first rectangular piece 151, the second rectangular piece 152, the third rectangular piece 153, the fourth rectangular piece 154, the fifth rectangular piece 155 and the sixth rectangular piece 156 are not connected to each other.

[0163] The third lag zone of the sixth rectangular strip 156 is connected to a universal lateral flow test strip 117 for detecting biotin and FITC-labeled analytes;

[0164] The nucleic acid signal amplification module and the signal display and detection module cover the outer surface of the bottom wall 102 of the elliptical cylinder; and expose the polymer waterproof and breathable material layer 148 of the virus attachment strip at the rectangular hole on the bottom wall of the elliptical cylinder.

[0165] The rapid diffusion material layer is made of filter paper, and the thickness of the rapid diffusion material layer is 0.02 mm.

[0166] The absorbent and breathable material layer is made of nylon fiber cloth; the thickness of the absorbent and breathable material layer is 0.01mm.

[0167] The polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the waterproof and breathable material layer is 0.01 mm.

[0168] The second type of embeddable micro-device for detecting target nucleic acids in exhaled air has a first base membrane 105, a second base membrane 107, a third base membrane 109, a fourth base membrane 111, a fifth base membrane 113, and a sixth base membrane 115 with a thickness of 20 μm and is made of butyl waterproof self-adhesive aluminum foil tape.

[0169] Example 21

[0170] The second type is a miniature device that can be embedded in a mask for detecting target nucleic acids in exhaled breath. Figure 6 and Figure 7 It includes a sample enrichment module, a nucleic acid signal amplification module, and a signal display and detection module;

[0171] The sample enrichment module includes an elliptical funnel-shaped cylinder 101, the small end of which is connected to the edge of the bottom wall 102 of the elliptical cylinder, and a rectangular hole 118 is provided on the bottom wall of the elliptical cylinder.

[0172] The nucleic acid signal amplification module (see Figure 8 , Figure 9 and Figure 10 )include

[0173] Virus attachment strip 104, the virus attachment strip includes a fast-diffusion material layer 149, a water-absorbing and breathable material layer 147 is provided in the middle of the upper surface of the fast-diffusion material layer 149, a polymer waterproof and breathable material layer 148 is provided in the middle of the lower surface of the fast-diffusion material layer 149, and a solution storage bubble 103 is connected to one end of the virus attachment strip.

[0174] The solution storage bubble is composed of a medical polyvinyl chloride bubble body connected to a medical aluminum foil substrate. The storage solution of Example 3 and the needle 150 are placed inside the solution storage bubble.

[0175] The first rectangular sheet 151 includes a first base film 105, and a circular hole with a diameter of 0.75 cm is punched in the middle of the first base film, which is the first circular hole 106. The edge of the first circular substrate 140 containing 40 μL of nucleic acid lysis buffer from Example 6, with a diameter of 0.8 cm, is attached to the edge of the first circular hole and dried to form a nucleic acid lysis zone.

[0176] The second rectangular sheet 152 includes a second base film 107, with a circular hole of 0.7 cm in diameter punched in the middle of the second base film, which is called the second circular hole 108. The second base film with the second circular hole is placed on a hydrophobic material, and 20 μL of the hydrophilic polymer aqueous solution of Example 9 is dropped into the second circular hole. After drying, a first dense film 141 is formed in the circular hole and at the edge of the circular hole, which is the first lag region.

[0177] The third rectangular sheet 153 includes a third base membrane 109, with a 0.75 cm diameter circular hole punched in the middle of the third base membrane, which is called the third circular hole 110. The edge of the second circular substrate 142 containing 40 μL of the amplification reagent of Example 12 with a diameter of 0.8 cm is attached to the edge of the third circular hole. After drying, it is a nucleic acid amplification area.

[0178] The fourth rectangular sheet 154 includes a fourth base film 111, with a circular hole of 0.7 cm in diameter punched in the middle of the fourth base film, which is the fourth circular hole 112. The fourth base film with the fourth circular hole is placed on a hydrophobic material, and 20 μL of the hydrophilic polymer aqueous solution of Example 9 is dropped into the fourth circular hole. After drying, a second dense film 143 is formed in the circular hole and at the edge of the circular hole, which is the second lag region.

[0179] The fifth rectangular sheet 155 includes a fifth base film 113, with a circular hole of 0.75 cm in diameter punched in the middle of the fifth base film, which is the fifth circular hole 114. The edge of the third circular substrate 144 containing 40 μL of the cutting reagent of Example 15 with a diameter of 0.8 cm is attached to the edge of the fifth circular hole and dried to form the probe cutting area.

[0180] The sixth rectangular sheet 156 includes a sixth base film 115, with a circular hole of 0.7 cm in diameter punched in the middle of the sixth base film, which is the sixth circular hole 116. The sixth base film with the sixth circular hole is placed on a hydrophobic material, and 20 μL of the hydrophilic polymer aqueous solution of Example 9 is dropped into the sixth circular hole. After drying, a third dense film 145 is formed inside the circular hole and at the edge of the circular hole, which is the third lag region.

[0181] The signal display detection module is a universal lateral flow test strip 117 for detecting biotin and FITC-labeled analytes;

[0182] The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular piece 151, and the first rectangular piece 151, the second rectangular piece 152, the third rectangular piece 153, the fourth rectangular piece 154, the fifth rectangular piece 155 and the sixth rectangular piece 156 are interconnected.

[0183] The third lag zone of the sixth rectangular strip 156 is connected to a universal lateral flow test strip 117 for detecting biotin and FITC-labeled analytes;

[0184] The nucleic acid signal amplification module and the signal display and detection module cover the outer surface of the bottom wall 102 of the elliptical cylinder; and expose the polymer waterproof and breathable material layer 148 of the virus attachment strip at the rectangular hole on the bottom wall of the elliptical cylinder.

[0185] The rapid diffusion material layer is made of cotton cloth and has a thickness of 0.5 mm.

[0186] The absorbent and breathable material layer is made of polyester fiber cloth; the thickness of the absorbent and breathable material layer is 0.5mm.

[0187] The polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the waterproof and breathable material layer is 0.5mm.

[0188] The second type of embeddable mask-style micro-device for detecting target nucleic acids in exhaled air has a first base membrane 105, a second base membrane 107, a third base membrane 109, a fourth base membrane 111, a fifth base membrane 113, and a sixth base membrane 115 with a thickness of 90 μm and is made of resin pressure-sensitive double-sided adhesive tape.

[0189] Example 22

[0190] Simulated detection of target nucleic acids in exhaled breath using an embeddable micro-device in a mask.

[0191] (1) The two sides of the elliptical funnel-shaped cylinder of the micro-device for detecting target nucleic acid in exhaled gas in Example 16 are fixed to the middle position inside the mask by hot pressing, so that the solution storage bubble is located directly above and the nucleic acid signal amplification module is located below.

[0192] (2) Connect an aerosol generator to the trachea of ​​the dummy and spray an aerosol containing the RNA sequence of the SARS-CoV-2 virus S protein (SEQ ID No. 5) into the elliptical funnel-shaped cylinder of the sample enrichment module of the above device (virus concentration: 1 copy / μL, solvent is enzyme-free water). After wearing a mask over the mouth and nose of the dummy for 10 minutes, press the solution storage bubble and puncture the medical aluminum foil with a needle to allow the storage solution to flow out. Place the dummy in a 37°C environment (simulating human body temperature) and continue wearing the mask for 30 minutes. Observe the test results using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. See Figure 11 (The right-hand strip in A).

[0193] Next, an aerosol containing the RNA sequence (SEQ ID No. 6) of the negative control HCoV-229E viral S protein (virus concentration: 1 copy / μL, solvent: enzyme-free water) was subjected to the above steps. The test results were observed using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. (See attached image.) Figure 11 (The left strip in A).

[0194] Example 23

[0195] Simulated detection of target nucleic acids in exhaled breath using an embeddable micro-device in a mask.

[0196] (1) The two sides of the elliptical funnel-shaped cylinder of the micro-device for detecting target nucleic acid in exhaled gas in Example 17 are fixed to the middle position inside the mask by hot pressing, so that the solution storage bubble is located directly above and the nucleic acid signal amplification module is located below.

[0197] (2) Connect an aerosol generator to the trachea of ​​the dummy and spray an aerosol containing the RNA sequence of matrix protein 2 of influenza A virus (SEQ ID No. 11) into the elliptical funnel-shaped cylinder of the sample enrichment module of the above device (virus concentration: 1 copy / μL, solvent is enzyme-free water). After wearing a mask over the mouth and nose of the dummy for 10 minutes, press the solution storage bubble and puncture the medical aluminum foil with a needle to allow the storage solution to flow out. Place the dummy in a 37°C environment (simulating human body temperature) and continue wearing the mask for 30 minutes. Observe the test results using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. See Figure 11 (The right-hand strip in B).

[0198] Next, an aerosol containing the RNA sequence of matrix protein 2 (SEQ ID No. 12) of the negative control influenza B virus (virus concentration: 1 copy / μL, solvent: enzyme-free water) was prepared using the above steps. The results were observed using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. (See attached image.) Figure 11 (The left strip in B).

[0199] Example 24

[0200] Simulated detection of target nucleic acids in exhaled breath using an embeddable micro-device in a mask.

[0201] (1) The two sides of the elliptical funnel-shaped cylinder of the micro-device for detecting target nucleic acid in exhaled gas in Example 18 are fixed to the middle position inside the mask by hot pressing, so that the solution storage bubble is located directly above and the nucleic acid signal amplification module is located below.

[0202] (2) Connect an aerosol generator to the trachea of ​​the dummy and spray an aerosol containing the RNA sequence of matrix protein 2 of influenza B virus (SEQ ID No. 12) into the elliptical funnel-shaped cylinder of the sample enrichment module of the above device (virus concentration: 1 copy / μL, solvent is enzyme-free water). After wearing a mask over the mouth and nose of the dummy for 10 minutes, press the solution storage bubble and puncture the medical aluminum foil with a needle to allow the storage solution to flow out. Place the dummy in a 37°C environment (simulating human body temperature) and continue wearing the mask for 30 minutes. Observe the test results using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. See Figure 11 (The right-hand strip in C).

[0203] Next, an aerosol containing the RNA sequence of matrix protein 2 (SEQ ID No. 11) of the negative control influenza A virus (virus concentration: 1 copy / μL, solvent: enzyme-free water) was prepared using the above steps. The results were observed using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. (See attached image.) Figure 11 (The left strip in C).

[0204] Example 25

[0205] Simulated detection of target nucleic acids in exhaled breath using an embeddable micro-device in a mask.

[0206] (1) The two sides of the elliptical funnel-shaped cylinder of the micro-device for detecting target nucleic acid in exhaled gas in Example 19 are fixed to the middle position inside the mask by hot pressing, so that the solution storage bubble is located directly above and the nucleic acid signal amplification module is located below.

[0207] (2) Connect an aerosol generator to the trachea of ​​the dummy and spray an aerosol containing the RNA sequence of the SARS-CoV-2 virus S protein (SEQ ID No. 5) into the elliptical funnel-shaped cylinder of the sample enrichment module of the above device (virus concentration: 1 copy / μL, solvent is enzyme-free water). After wearing a mask over the mouth and nose of the dummy for 10 minutes, press the solution storage bubble and puncture the medical aluminum foil with a needle to allow the storage solution to flow out. Place the dummy in a 37°C environment (simulating human body temperature) and continue wearing the mask for 30 minutes. Observe the test results using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. See Figure 12 (The right-hand strip in A).

[0208] An aerosol containing the RNA sequence (SEQ ID No. 6) of the negative control HCoV-229E viral S protein (virus concentration: 1 copy / μL, solvent: enzyme-free water) was prepared using the above steps. The results were observed using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. (See attached image). Figure 12 (The left strip in A).

[0209] Example 26

[0210] Simulated detection of target nucleic acids in exhaled breath using an embeddable micro-device in a mask.

[0211] (1) The two sides of the elliptical funnel-shaped cylinder of the micro-device for detecting target nucleic acid in exhaled gas in Example 20 are fixed to the middle position inside the mask by hot pressing, so that the solution storage bubble is located directly above and the nucleic acid signal amplification module is located below.

[0212] (2) Connect an aerosol generator to the trachea of ​​the dummy and spray an aerosol containing the RNA sequence of matrix protein 2 of influenza A virus (SEQ ID No. 11) into the elliptical funnel-shaped cylinder of the sample enrichment module of the above device (virus concentration: 1 copy / μL, solvent is enzyme-free water). After wearing a mask over the mouth and nose of the dummy for 10 minutes, press the solution storage bubble and puncture the medical aluminum foil with a needle to allow the storage solution to flow out. Place the dummy in a 37°C environment (simulating human body temperature) and continue wearing the mask for 30 minutes. Observe the test results using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. See Figure 12 (The right-hand strip in B).

[0213] An aerosol containing the RNA sequence of matrix protein 2 (SEQ ID No. 12) of the negative control influenza B virus (virus concentration: 1 copy / μL, solvent: enzyme-free water) was prepared using the above steps. The results were observed using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. (See attached image.) Figure 12 (The left strip in B).

[0214] Example 27

[0215] Simulated detection of target nucleic acids in exhaled breath using an embeddable micro-device in a mask.

[0216] (1) The two sides of the elliptical funnel-shaped cylinder of the micro-device for detecting target nucleic acid in exhaled gas in Example 21 are fixed to the middle position inside the mask by hot pressing, so that the solution storage bubble is located directly above and the nucleic acid signal amplification module is located below.

[0217] (2) Connect an aerosol generator to the trachea of ​​the dummy and spray an aerosol containing the RNA sequence of matrix protein 2 of influenza B virus (SEQ ID No. 12) into the elliptical funnel-shaped cylinder of the sample enrichment module of the above device (virus concentration: 1 copy / μL, solvent is enzyme-free water). After wearing a mask over the mouth and nose of the dummy for 10 minutes, press the solution storage bubble and puncture the medical aluminum foil with a needle to allow the storage solution to flow out. Place the dummy in a 37°C environment (simulating human body temperature) and continue wearing the mask for 30 minutes. Observe the test results using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. See Figure 12 (The right-hand strip in C).

[0218] An aerosol containing the RNA sequence of matrix protein 2 (SEQ ID No. 11) of the negative control influenza A virus (virus concentration: 1 copy / μL, solvent: enzyme-free water) was prepared using the above steps. The results were observed using a universal lateral flow test strip for detecting biotin and FITC-labeled analytes. (See attached image). Figure 12 (The left strip in C).

[0219] The technical solutions disclosed and proposed in this invention can be implemented by those skilled in the art by appropriately modifying the conditions and routes, etc. Although the methods and preparation techniques of this invention have been described through preferred embodiments, those skilled in the art can obviously modify or recombine the methods and technical routes described herein without departing from the content, spirit, and scope of this invention to achieve the final preparation technique. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included within the spirit, scope, and content of this invention.

Claims

1. An embeddable micro-device for detecting target nucleic acids in exhaled breath, including a sample enrichment module, characterized in that: It also includes a nucleic acid signal amplification module and a signal display and detection module; the sample enrichment module includes an elliptical funnel-shaped cylinder (1), the small end of which is connected to the edge of the bottom wall (2) of the elliptical cylinder, and a rectangular hole (18) is provided on the bottom wall of the elliptical cylinder; the nucleic acid signal amplification module includes a virus attachment strip (4), the virus attachment strip includes a rapid diffusion material layer (49), a water-absorbing and breathable material layer (47) is provided in the middle of the upper surface of the rapid diffusion material layer (49), and a polymer waterproof and breathable material is provided in the middle of the lower surface of the rapid diffusion material layer (49). The material layer (48) has a solution storage bubble (3) connected to one end of the virus attachment strip; the solution storage bubble is composed of medical polyvinyl chloride as the bubble body and medical aluminum foil as the substrate, and a storage solution and a needle (50) are set inside the solution storage bubble; the first rectangular sheet (51) includes a first base film (5), and a circle with a diameter of 0.35-0.8cm is drawn in the middle of the first base film, which is the first circle (6). The outside of the first circle is sealed with wax as a hydrophobic area, and 3-40μL of nucleic acid lysis buffer is set inside the first circle. After drying, it is the nucleic acid lysis area; the second rectangular sheet (52) includes a second base film (7), and a circle with a diameter of 0.35-0.8cm is drawn in the middle of the second base film. A circle with a diameter of 0.2-0.7 cm is the second circle (8). The outer part of the second circle is sealed with wax as a hydrophobic region. 4-20 μL of hydrophilic polymer aqueous solution is placed inside the second circle. After drying, it is the first lag region. The third rectangular sheet (53) includes the third base membrane (9). A circle with a diameter of 0.35-0.8 cm is drawn in the middle of the third base membrane, which is the third circle (10). The outer part of the third circle is sealed with wax as a hydrophobic region. 3-40 μL of amplification reagent is placed inside the third circle. After drying, it is the nucleic acid amplification region. The fourth rectangular sheet (54) includes the fourth base membrane (11). A circle with a diameter of 0.2-0.7 cm is drawn in the middle of the fourth base membrane. A circle with a diameter of -0.7cm is the fourth circle (12). The area outside the fourth circle is sealed with wax as a hydrophobic region. 4-20μL of hydrophilic polymer aqueous solution is placed inside the fourth circle. After drying, it becomes the second lag region. The fifth rectangular sheet (55) includes the fifth base film (13). A circle with a diameter of 0.35-0.8cm is drawn in the middle of the fifth base film, which is the fifth circle (14). The area outside the fifth circle is sealed with wax as a hydrophobic region. 3-40μL of cutting reagent is placed inside the fifth circle. After drying, it becomes the probe cutting region. The sixth rectangular sheet (56) includes the sixth base film (15). A circle with a diameter of 0.2-0.7cm is drawn in the middle of the sixth base film.A 7cm circle is the sixth circle (16). The area outside the sixth circle is sealed with wax as a hydrophobic region. A 4-20μL aqueous solution of hydrophilic polymer is placed inside the sixth circle. After drying, it forms the third lag region. The signal display detection module is a universal lateral flow test strip (17) for detecting biotin and FITC-labeled analytes. The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular sheet (51). The first rectangular sheet (51), the second rectangular sheet (52), the third rectangular sheet (53), the fourth rectangular sheet (54), the fifth rectangular sheet (55), and the sixth rectangular sheet (56) are connected to each other or not connected. The third lag region of the sixth rectangular sheet (56) is connected to the universal lateral flow test strip (17) for detecting biotin and FITC-labeled analytes. The nucleic acid signal amplification module and the signal display detection module cover the outer surface of the bottom wall (2) of the elliptical cylinder. The polymer waterproof and breathable material layer (48) of the virus attachment strip is exposed at the rectangular hole on the bottom wall of the elliptical cylinder. The storage solution consists of 1–5 T / mL of ribonuclease inhibitor, 0.01%–0.05% TritiumX-100, 0.5%–2.5% CHAPS, and 0.01%–0.05% NP-40, with the remainder being nuclease-free water. The rapid diffusion material layer is made of cotton cloth or filter paper, and the thickness of the rapid diffusion material layer is 0.02-0.5 mm. The absorbent and breathable material layer is made of cellulose paper, nylon fiber cloth, or polyester fiber cloth; the thickness of the absorbent and breathable material layer is 0.01–0.5 mm. The polymer waterproof and breathable material layer is made of meltblown fabric; the thickness of the polymer waterproof and breathable material layer is 0.01-0.5 mm; The aqueous solution of the hydrophilic polymer has a concentration of 4wt%-20wt%; the hydrophilic polymer is polyvinyl alcohol with a weight average molecular weight of 30,000-100,000, polyethylene glycol with a weight average molecular weight of 50,000-150,000, polypropylene glycol with a weight average molecular weight of 50,000-150,000, sodium carboxymethyl cellulose with a weight average molecular weight of 30,000-250,000, or sodium alginate with a weight average molecular weight of 10,000-20,000.

2. An embeddable micro-device for detecting target nucleic acids in exhaled breath, including a sample enrichment module, characterized in that: It also includes a nucleic acid signal amplification module and a signal display and detection module; the sample enrichment module includes an elliptical funnel-shaped cylinder (101), the small end of which is connected to the edge of the bottom wall (102) of the elliptical cylinder, and a rectangular hole (118) is provided on the bottom wall of the elliptical cylinder; the nucleic acid signal amplification module includes a virus attachment strip (104), which includes a rapid diffusion material layer (149), a water-absorbing and breathable material layer (147) is provided in the middle of the upper surface of the rapid diffusion material layer (149), and a polymer waterproof and breathable material layer (148) is provided in the middle of the lower surface of the rapid diffusion material layer (149); a solution storage bubble (103) is connected to one end of the virus attachment strip; the solution storage bubble (103) is connected to the virus attachment strip. The storage bubble is constructed by connecting a medical-grade polyvinyl chloride (PVC) bubble body with a medical-grade aluminum foil substrate. A storage solution and a needle (150) are placed inside the storage bubble. A first rectangular sheet (151) includes a first base membrane (105), with a circular hole (106) of 0.3-0.75 cm in diameter punched in the center. A first circular substrate (140) containing 3-40 μL of nucleic acid lysis buffer (0.35-0.8 cm in diameter) is attached to the edge of the first circular hole and dried, forming a nucleic acid lysis zone. A second rectangular sheet (152) includes a second base membrane (107), with a circular hole (108) of 0.2-0.7 cm in diameter punched in the center. The second base membrane with the second circular hole is placed in a porous... On the aqueous material, 4-20 μL of a hydrophilic polymer aqueous solution is dropped into the second circular hole. After drying, a first dense membrane (141) is formed inside and at the edge of the circular hole, which is the first lag region. The third rectangular sheet (153) includes a third base membrane (109). A circular hole with a diameter of 0.3-0.75 cm is punched in the middle of the third base membrane, which is the third circular hole (110). The edge of the second circular substrate (142) with a diameter of 0.35-0.8 cm containing 3-40 μL of amplification reagent is attached to the edge of the third circular hole. After drying, it is the nucleic acid amplification region. The fourth rectangular sheet (154) includes a fourth base membrane (111). A circular hole with a diameter of 0.2-0.7 cm is punched in the middle of the fourth base membrane, which is the fourth circular hole (112). The fourth base film with the fourth circular hole is placed on a hydrophobic material, and 4-20 μL of hydrophilic polymer aqueous solution is dropped into the fourth circular hole. After drying, a second dense film (143) is formed inside and at the edge of the circular hole, which is the second lag region. The fifth rectangular sheet (155) includes the fifth base film (113), and a circular hole with a diameter of 0.3-0.75 cm is punched in the middle of the fifth base film, which is the fifth circular hole (114). The edge of the third circular substrate (144) with a diameter of 0.35-0.8 cm containing 3-40 μL of cutting reagent is attached to the edge of the fifth circular hole. After drying, it is the probe cutting region. The sixth rectangular sheet (156) includes the sixth base film (115), and a circular hole with a diameter of 0.2-0.8 cm is punched in the middle of the sixth base film.A 7cm circular hole is designated as the sixth circular hole (116). The sixth base membrane with the sixth circular hole is placed on a hydrophobic material, and 4-20μL of a hydrophilic polymer aqueous solution is added to the sixth circular hole. After drying, a third dense membrane (145) is formed inside and at the edge of the circular hole, which is the third lag region. The signal display detection module is a universal lateral flow test strip (117) for detecting biotin and FITC-labeled analytes. The other end of the virus attachment strip is connected to the nucleic acid lysis region of the first rectangular sheet (151). The first rectangular sheet (151) and the second rectangular sheet (151) are connected to the nucleic acid lysis region of the first rectangular sheet (151). The first rectangular strip (152), the third rectangular strip (153), the fourth rectangular strip (154), the fifth rectangular strip (155), and the sixth rectangular strip (156) are connected or not connected; the third lag zone of the sixth rectangular strip (156) is connected to a universal lateral flow test strip (117) for detecting biotin and FITC-labeled analytes; the nucleic acid signal amplification module and the signal display detection module cover the outer surface of the bottom wall (102) of the elliptical cylinder; and the polymer waterproof and breathable material layer (148) of the virus attachment strip is exposed at the rectangular holes on the bottom wall of the elliptical cylinder.

3. The apparatus according to claim 1 or 2, characterized in that: The nucleic acid lysis buffer consists of: 0.001%–0.05% NP-40, 1%–3% CHAPS, 0.5%–3% lysozyme, 2%–10% sucrose, and the remainder is 10–100 mM Tris buffer at pH 7.

5.

4. The apparatus according to claim 1 or 2, characterized in that: The amplification reagents are: TvsX 50–300 ng / μL, TvsY 30–200 ng / μL, T4gp3 2500–2000 ng / μL, polymerase 20–150 ng / μL, creatine kinase 50–450 ng / μL, RNase inhibitor 0.02–50 ng / μL, AMV reverse transcriptase 0.02–50 ng / μL, RNase H 0.2–80 ng / μL, and Tris buffer 20. ~100mM, potassium acetate 70-120mM, magnesium acetate 10-20mM, dithiothreitol 1-4mM, PEG 4wt%-10wt%, dNTP 300-800μM, ATP 2-6mM, creatine phosphate 30-80mM, BSA 0.01wt%-5wt%, primers 1-10μM, wherein the polymerase is Bst polymerase or Sat polymerase; and the PEG is PEG20000 or PEG35000.

5. The apparatus according to claim 1 or 2, characterized in that: The cleavage reagent consists of: 3–20 mM magnesium salt, 0.01 wt%–5% BSA, 40–2000 ng / μL Cas12a, 1–10 mM DTT, 0.2–1 pmol FB probe, and 0.05–3 μM viral gene targeting gRNA; the balance is 3–50 mM Tris-HCl buffer at pH 7.0–8.5; the magnesium salt is magnesium chloride or magnesium acetate; wherein the FB probe is a single-stranded nucleic acid sequence labeled with FITC fluorescent pigment at the 5' end and biotin at the 3' end.

6. The apparatus according to claim 1, characterized in that: The thickness of the first base film (5), the second base film (7), the third base film (9), the fourth base film (11), the fifth base film (13), and the sixth base film (15) is 0.01 to 0.5 mm, and the material is cellulose paper, cotton cloth, nylon fiber cloth or polyester fiber cloth.

7. The apparatus according to claim 2, characterized in that: The thickness of the first base film (105), the second base film (107), the third base film (109), the fourth base film (111), the fifth base film (113), and the sixth base film (115) is 20-90 μm, and the material is biaxially oriented polypropylene transparent tape, butyl waterproof self-adhesive aluminum foil tape, or resin pressure-sensitive double-sided tape.

8. The apparatus according to claim 1, characterized in that: The wax is microcrystalline wax, paraffin wax, petrolatum, or polypropylene.

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