Y-shaped DNA probe functionalized field effect transistor sensor for detecting novel coronavirus nucleic acid and its preparation method and application

Abstract

The present invention discloses a Y-shaped DNA probe functionalized field effect transistor sensor for detecting novel coronavirus nucleic acid, and its preparation method and application. The Y-shaped DNA probe is formed by hybridization of oligonucleotide chain S1 and oligonucleotide chain S2 through base complementary pairing; the base sequence of oligonucleotide chain S1 is such as SEQ ID NO.1; the base sequence of oligonucleotide chain S2 is such as SEQ ID NO.2. The field effect transistor sensor includes: a field effect transistor sensor device and the DNA probe, the field effect transistor device is provided with a semiconductor layer, and the semiconductor layer is modified and fixed with the Y-shaped DNA probe. Rapid and highly sensitive detection of low viral load samples is achieved. This method does not require the extraction and amplification of novel coronavirus nucleic acid, simplifies the detection process, the average detection time is less than 5 minutes, and the detection limit is 10 ‑16 mol / L.

Description

Technical Field

[0001] The present invention relates to the field of biological detection technology, and in particular to a Y-shaped DNA probe-functionalized field-effect transistor sensor for detecting novel coronavirus nucleic acid, and a preparation method and application thereof. Background Art

[0002] Since the initial outbreak of COVID-19 in 2019, rapid and highly sensitive detection of SARS-CoV-2 has become a key focus of epidemic prevention and control. Nucleic acid testing is currently the most widely used method for detecting SARS-CoV-2. Real-time fluorescence quantitative reverse transcription nucleic acid amplification (RT-PCR) technology has become the gold standard for SARS-CoV-2 detection due to its high sensitivity and strong specificity. However, RT-PCR requires the extraction and amplification of SARS-CoV-2 nucleic acid, relies on specialized equipment, and typically takes more than 2 hours to detect, making it unsuitable for immediate detection and large-scale screening of SARS-CoV-2.

[0003] Therefore, developing a convenient method for rapid and highly sensitive detection of SARS-CoV-2 is of great significance for epidemic prevention and control and large-scale screening of SARS-CoV-2. Summary of the Invention

[0004] The purpose of the present invention is to provide a Y-shaped DNA probe functionalized field-effect transistor sensor for detecting novel coronavirus nucleic acid, its preparation method and application, which can achieve rapid and highly sensitive detection of low viral load samples. This method does not require the extraction and amplification of SARS-CoV-2 nucleic acid, simplifies the detection process, and has an average detection time of less than 5 minutes and a detection limit of 10 -16 mol / L.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] In the first aspect of the present invention, a Y-shaped DNA probe for detecting novel coronavirus nucleic acid is provided, wherein the Y-shaped DNA probe is formed by hybridization of oligonucleotide chain S1 and oligonucleotide chain S2 through complementary base pairing;

[0007] The base sequence of the oligonucleotide chain S1 is: 5'-TTTTTTACCTAGCTCCGCAGACGG-3'; the base sequence of the oligonucleotide chain S2 is: 5'-CCATAACCTTTCCACATAAGCTAGGTAAAA-3'.

[0008] Furthermore, the Y-shaped DNA probe also includes: at least one of the bases in the oligonucleotide chain S1 and the oligonucleotide chain S2 is modified, and the modification includes at least one of phosphorylation, methylation, amination, sulfhydrylation, replacement of oxygen with sulfur, replacement of oxygen with selenium or isotopization; or at least one of the bases in the oligonucleotide chain S1 and the oligonucleotide chain S2 is connected to at least one of a fluorescent marker, biotin, digoxigenin, and a nanoluminescent material.

[0009] In the second aspect of the present invention, a field effect transistor sensor for detecting novel coronavirus nucleic acid is provided, comprising: a field effect transistor sensor device and the Y-shaped DNA probe, wherein the field effect transistor device is provided with a semiconductor layer, and the semiconductor layer is modified and fixed with the Y-shaped DNA probe.

[0010] Furthermore, the field effect transistor device includes an insulating substrate, a semiconductor layer and electrodes provided on the insulating substrate, wherein the semiconductor layer is modified and fixed with the Y-shaped DNA probe.

[0011] In a third aspect of the present invention, a method for preparing a field effect transistor sensor for detecting novel coronavirus nucleic acid is provided, the method comprising:

[0012] obtaining a field effect transistor device;

[0013] The field effect transistor device is placed in an ethanol solution of 3-aminopropyltriethoxysilane (APTES) and then heated to form an APTES-modified FET;

[0014] Immersing the APTES-modified FET in a sodium salt solution of sulfosuccinimidyl cyclohexane-1-carboxylate (sulfo-SMCC) to obtain a sulfo-SMCC-functionalized FET;

[0015] The connecting molecule APTES and the sulfo-SMCC functionalized FET were immersed in a solution of oligonucleotide chain S1 and incubated, and then rinsed and immersed in a solution of oligonucleotide chain S2 to obtain a field effect transistor sensor for detecting novel coronavirus nucleic acid.

[0016] Furthermore, the molar fraction of the ethanol solution of 3-aminopropyltriethoxysilane (APTES) is 5% to 8%.

[0017] Furthermore, the field effect transistor device is placed in an ethanol solution of 3-aminopropyltriethoxysilane (APTES) and incubated for 50 to 60 minutes, and the heating condition is 105° C. to 115° C. for 25 to 35 minutes.

[0018] Furthermore, the concentration of the sulfosuccinimidyl cyclohexane-1-carboxylate sodium salt (sulfo-SMCC) is 1 to 3 mg / mL.

[0019] Furthermore, the APTES-modified FET is immersed in a sulfo-succinimidyl cyclohexane-1-carboxylate sodium salt (sulfo-SMCC) solution and incubated for 1.5 to 2 hours.

[0020] Furthermore, the linker molecule APTES and the sulfo-SMCC functionalized FET are immersed in the solution of oligonucleotide chain S1 and incubated for 10 to 12 hours, and then rinsed and immersed in the solution of oligonucleotide chain S2 and incubated for 0.5 to 1 hour.

[0021] In the fourth aspect of the present invention, the application of the Y-shaped DNA probe or the field effect transistor sensor in the preparation of a novel coronavirus nucleic acid detection product is provided.

[0022] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0023] 1. The present invention provides a Y-shaped DNA probe functionalized field effect transistor sensor for detecting novel coronavirus nucleic acid, as well as its preparation method and application. By organically combining the Y-shaped DNA probe with FET, the stability and sensitivity of SARS-CoV-2 detection are effectively improved. Compared with RT-PCR technology, the sensor does not require extraction and amplification of SARS-CoV-2 nucleic acid, and can achieve rapid and highly sensitive detection of SARS-CoV-2. The detection time is less than 5 minutes, and the detection limit is 10 -16 mol / L.

[0024] 2. The field-effect transistor sensor (YDN FETs) for detecting novel coronavirus nucleic acid in the present invention has the advantages of fast response speed, high sensitivity, easy miniaturization and integration, and is expected to be used to construct a portable biosensor for instant detection of SARS-CoV-2, effectively shortening the SARS-CoV-2 detection time and reducing the detection cost, realizing large-scale screening of the virus, and providing new ideas for family self-examination, health monitoring and management. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following is a brief introduction to the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0026] Figure 1This is a flow chart for preparing YDN FETs in Example 1 of the present invention.

[0027] Figure 2 This is a graph showing the specific response of YDN FETs to SARS-CoV-2 in Example 2 of the present invention.

[0028] Figure 3 This is a sensor response diagram of the YDN FETs to the sample to be tested in Example 3 of the present invention. DETAILED DESCRIPTION

[0029] The present invention will be described in detail below in conjunction with specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific embodiments and examples are for illustrating the present invention, rather than for limiting the present invention.

[0030] Throughout this specification, unless otherwise specified, the terms used herein should be understood as having the same meaning as commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In the event of any conflict, the present specification shall take precedence.

[0031] Unless otherwise specified, all raw materials, reagents, instruments, and equipment used in the present invention can be purchased from the market or obtained through existing methods. Steps S1, S2, S3, etc. of the present invention do not represent a strict order relationship and can be adjusted as needed.

[0032] The technical solution of the embodiment of the present application is to solve the above technical problems, and the overall idea is as follows:

[0033] According to a typical embodiment of the present invention, a Y-shaped DNA probe for detecting novel coronavirus nucleic acid is provided, wherein the Y-shaped DNA probe is formed by hybridization of oligonucleotide chain S1 and oligonucleotide chain S2 through complementary base pairing;

[0034] The base sequence of the oligonucleotide chain S1 is: 5'-TTTTTTACCTAGCTCCGCAGACGG-3' (SEQ ID NO. 1); the base sequence of the oligonucleotide chain S2 is: 5'-CCATAACCTTTCCACATAAGCTAGGTAAAA-3' (SEQ ID NO. 2).

[0035] The Y-shaped DNA probe for detecting novel coronavirus nucleic acid specifically targets the SARS-CoV-2 ORF1ab nucleic acid sequence. The target molecule sequence of the Y-shaped DNA probe for detecting novel coronavirus nucleic acid is: 5'-CCGTCTGCGGTATGTGG-AAAGGTTATGG-3'. This Y-shaped DNA probe overcomes the shortcomings of traditional single-stranded DNA probes, which are prone to aggregation and non-covalent adsorption, effectively improving the recognition ability of target molecules and enhancing the stability and sensitivity of the sensor.

[0036] Preferably, the Y-shaped DNA probe further comprises oligonucleotides formed by adding, reducing or replacing one or more bases in the base sequence of the oligonucleotide chain S1 and the oligonucleotide chain S2.

[0037] Preferably, the Y-shaped DNA probe further includes: at least one of the bases in the oligonucleotide chain S1 and the oligonucleotide chain S2 is modified, and the modification includes at least one of phosphorylation, methylation, amination, sulfhydrylation, replacement of oxygen with sulfur, replacement of oxygen with selenium or isotopization; or at least one of the bases in the oligonucleotide chain S1 and the oligonucleotide chain S2 is connected to at least one of a fluorescent marker, biotin, digoxigenin, and a nanoluminescent material.

[0038] As a specific embodiment, the base sequence of the oligonucleotide chain S1 is: 5'-SH-TTTTTTACCTAGCTCCGCAGACGG-3'; the base sequence of the oligonucleotide chain S2 is: 5'-CCATAACCTTTCCACATAAGCTAGGTAAAA-3', wherein SH represents sulfhydrylation.

[0039] According to another typical embodiment of the present invention, a field effect transistor sensor for detecting novel coronavirus nucleic acid is provided, comprising: a field effect transistor sensor device and the Y-shaped DNA probe, wherein:

[0040] A semiconductor layer is provided on the field effect transistor device, and the Y-shaped DNA probe is modified and fixed on the semiconductor layer.

[0041] As a specific embodiment, the field effect transistor device includes an insulating substrate, a semiconductor layer and electrodes provided on the insulating substrate, and the semiconductor layer is modified and fixed with the Y-shaped DNA probe.

[0042] The semiconductor material of the semiconductor layer forms a conductive channel after being turned on.

[0043] The DNA probe is modified and fixed to the surface of the semiconductor layer by an adsorption method, a cross-linking method, a covalent bonding method, an embedding method or a biological tissue fixation method.

[0044] The insulating substrate is made of silicon dioxide, quartz, insulating glass, mica, polyethylene terephthalate film, polyimide film or polydimethylsiloxane film.

[0045] The semiconductor layer is graphene, oxide semiconductor, transition metal chalcogenide, silicon or germanium.

[0046] The electrode is a patterned electrode, the thickness of the electrode material is 20 to 2000 nanometers, and the electrode can be made of metal elements such as gold, silver, copper, titanium, chromium, as well as conductive silicide, carbide, conductive polymer, etc.

[0047] As a specific embodiment, the present invention prepares a field effect transistor device by ultraviolet photolithography, thermal evaporation, and magnetron sputtering. The specific operation steps may be: spin-coating a layer of positive photoresist on an insulating substrate, and then using an ultraviolet photolithography machine under the protection of a mask to carve an electrode channel on the crystal surface; using a thermal evaporation apparatus to grow a Cr / Au electrode on the electrode channel; under argon protection, using a magnetron sputtering method to grow a semiconductor material thin film on the electrode channel to prepare a FET (the specific preparation method of the field effect transistor device can be referred to patent: CN114354722A or reference: DOI: 10.1002 / adma.202203224).

[0048] According to a typical embodiment of the present invention, a method for preparing a Y-shaped DNA probe functionalized device for detecting novel coronavirus nucleic acid is provided, the method comprising:

[0049] Step S1, obtaining a field effect transistor device;

[0050] In step S1, the field effect transistor device may be processed as follows:

[0051] Polymethyl methacrylate (PMMA) was spin-coated on the device surface to passivate the source and drain electrodes, and then an electron beam lithography (EBL) system was used to expose the sensing area covered by PMMA.

[0052] Step S2, placing the field effect transistor device in an ethanol solution of 3-aminopropyltriethoxysilane (APTES) and incubating and then heating to obtain an APTES-modified FET;

[0053] In the step S2,

[0054] The reason for using APTES to modify FET is to introduce highly active functional groups on the insulating substrate, which can be used to fix biomacromolecules such as enzymes, antibodies, and nucleic acids.

[0055] The molar fraction of the ethanol solution of 3-aminopropyltriethoxysilane (APTES) is 5% to 8%, wherein the ethanol solution is a 95% ethanol solution. A lower molar fraction results in a lower density of introduced functional groups, while a higher molar fraction may lead to uneven modification.

[0056] The field effect transistor device is placed in an ethanol solution of 3-aminopropyltriethoxysilane (APTES) and incubated for 50 to 60 minutes. The heating condition is 105° C. to 115° C. for 25 to 35 minutes.

[0057] Step S3, immersing the APTES-modified FET in a sulfo-succinimidyl cyclohexane-1-carboxylate sodium salt (sulfo-SMCC) solution (sulfo-SMCC) for incubation to obtain a sulfo-SMCC-functionalized FET;

[0058] In the step S3,

[0059] The reason for functionalizing the APTES-modified FET with sulfo–SMCC is to facilitate the binding of the oligonucleotide chain S1 to the sensing region of the functionalized device via the covalent bond formed between the sulfhydryl group and sulfo–SMCC in the subsequent step;

[0060] The concentration of sulfosuccinimidyl cyclohexane-1-carboxylate sodium salt (sulfo-SMCC) is preferably 1-3 mg / mL. A concentration that is too low results in less bound nucleic acid, while a concentration that is too high results in inability to dissolve sulfo-SMCC.

[0061] The APTES-modified FET is immersed in a sulfo-succinimidyl cyclohexane-1-carboxylate sodium salt (sulfo-SMCC) solution and incubated for 1.5 to 2 hours.

[0062] Step S4: Immerse the linker molecule APTES and the sulfo-SMCC-functionalized FET in a solution of oligonucleotide chain S1 and incubate. After rinsing, the sensor is immersed in a solution of oligonucleotide chain S2 and incubated to obtain a field-effect transistor sensor for detecting novel coronavirus nucleic acid. In a specific embodiment, the linker molecule APTES is an ethanol solution of 3-aminopropyltriethoxysilane (APTES), with a molar fraction of 5% to 8% in ethanol. The ethanol solution is a 95% ethanol solution.

[0063] According to another typical embodiment of the present invention, the use of the Y-shaped DNA probe for detecting novel coronavirus nucleic acid in the preparation of novel coronavirus nucleic acid detection products is provided.

[0064] When the Y-shaped DNA probe fixed on the FET surface binds to the SARS-CoV-2 nucleic acid molecules in the test solution according to the principle of base complementary pairing, the charge density on the surface of the FET sensing area changes, thereby affecting the carrier density of the oxide channel material, and thus causing a change in the FET current response signal. By testing the current signals corresponding to SARS-CoV-2 nucleic acid solutions with different concentrations, a standard curve of the relationship between the current signal and the SARS-CoV-2 nucleic acid concentration was obtained (y = 0.10x + 1.83, R 2 =0.9974). By monitoring the current response of the FET to the sample solution to be tested, rapid and highly sensitive detection of SARS-CoV-2 nucleic acid was achieved.

[0065] The following is a detailed description of a Y-shaped DNA probe for detecting novel coronavirus nucleic acid, its preparation method, and application in combination with examples, comparative examples, and experimental data.

[0066] Example 1: Field Effect Transistor Sensor for Detecting Novel Coronavirus Nucleic Acid (YDN FETs) and Its Preparation Method

[0067] (1) Field effect transistor devices were prepared by ultraviolet lithography, thermal evaporation, and magnetron sputtering. PMMA was spin-coated on the device surface to passivate the source and drain electrodes. EBL was then used to expose the PMMA-covered sensing area to produce a FET.

[0068] (2) The FET was placed in a 5% molar fraction APTES ethanol solution and incubated for 60 min, and heated at 110°C for 30 min.

[0069] (3) The APTES-modified FET was immersed in a 2 mg / mL sulfo–SMCC solution and incubated for 2 h.

[0070] (4) The FET functionalized with the linker molecules APTES and sulfo–SMCC was immersed in a solution of oligonucleotide chain S1 and incubated for 12 h. The oligonucleotide chain S1 was combined with the functionalized device through the covalent bond formed between the thiol group and sulfo–SMCC.

[0071] (5) The device functionalized with oligonucleotide chain S1 was rinsed with PBS buffer and then immersed in a solution of oligonucleotide chain S2 for incubation for 1 hour. Oligonucleotide chain S1 and oligonucleotide chain S2 were combined through base complementary pairing to obtain YDN FETs. ( Figure 1 ).

[0072] Example 2: Specific detection of SARS-CoV-2 by YDN FETs

[0073] (1) Add 2 μL of PBS buffer solution to the sensing area of ​​the YDN FETs and set the test parameter source-drain voltage V d is 0.2V, gate voltage V g The transfer characteristic curve of PBS buffer solution was obtained in the range of -0.6 to 0.9 V, and the background current I was recorded. o .

[0074] (2) The concentration is 10 –12 2 μL of each 10 mol / L SARS-CoV-2 RNA solution, single-base mismatch, double-base mismatch, and triple-base mismatch nucleic acid solution was added dropwise to the sensing area of ​​the YDN FETs. The Y-shaped DNA probe was incubated for 5 minutes to allow binding of the nucleic acid. The sample was then rinsed three times with PBS buffer and reapplied with 2 μL of PBS buffer for testing.

[0075] (3) Set the test parameter source-drain voltage V d is 0.2V, gate voltage V g The range is -0.6 to 0.9 V, and the transfer characteristic curves of different nucleic acid solutions are recorded, and the response current I is recorded.

[0076] (4) Calculate the sensing response of different nucleic acid solutions, such as Figure 2 As shown, the current response of the SARS-CoV-2 RNA solution is significantly higher than that of the solutions with single-base mismatch, double-base mismatch, and triple-base mismatch, indicating that the prepared YDN FETs can achieve specific detection of SARS-CoV-2.

[0077] Example 3: Rapid and highly sensitive detection of SARS-CoV-2 using YDN FETs

[0078] 1. Add 2 μL of PBS buffer solution to the sensing area of ​​YDN FETs and set the test parameter source-drain voltage V d is 0.2V, gate voltage V g The transfer characteristic curve of PBS buffer solution was obtained in the range of -0.6 to 0.9 V, and the background current I was recorded. o .

[0079] 2. Configure gradient concentration (10 -18 ~10 -9 mol / L) SARS-CoV-2 nucleic acid solution, 2 μL of solutions with different concentrations were respectively added dropwise to the sensing area of ​​YDN FETs.

[0080] 3. After incubation for 5 minutes, rinse the device with PBS buffer solution, then add 2 μL of PBS buffer solution and set the test parameter source-drain voltage V d is 0.2V, gate voltage V gThe transfer characteristic curves of SARS-CoV-2 nucleic acid solutions with different concentrations were obtained in the range of -0.6 to 0.9 V, and the response current I was recorded.

[0081] 4. Calculate the sensor response ΔI / I at different concentrations of SARS-CoV-2 nucleic acid o , and obtain the corresponding relationship between the device sensing response and the SARS-CoV-2 nucleic acid concentration ( Figure 3 As shown in the figure, the linear detection range of YDN FETs for SARS-CoV-2 nucleic acid is 10 -16 ~10 -12 mol / L.

[0082] 5. Use YDN FETs to test unknown samples and calculate their sensor response ΔI / I o , when ΔI / I o When ΔI / I o When ΔI / I o If the value is between 10% and 15%, it is recommended to conduct a re-examination as a positive result.

[0083] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements, but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0084] Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0085] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A Y-shaped DNA probe for detecting novel coronavirus nucleic acid, characterized in that: The Y-shaped DNA probe is formed by hybridization of oligonucleotide chain S1 and oligonucleotide chain S2 through complementary base pairing; The base sequence of the oligonucleotide chain S1 is: 5'-TTTTTTACCTAGCTCCGCAGACGG-3'; the base sequence of the oligonucleotide chain S2 is: 5'-CCATAACCTTTCCACATAAGCTAGGTAAAA-3'.

2. A Y-shaped DNA probe for detecting novel coronavirus nucleic acid according to claim 1, characterized in that: The Y-shaped DNA probe also includes: at least one base in the oligonucleotide chain S1 and the oligonucleotide chain S2 is modified, and the modification includes at least one of phosphorylation, methylation, amination, sulfhydrylation, replacement of oxygen with sulfur, replacement of oxygen with selenium or isotopization; or at least one base in the oligonucleotide chain S1 and the oligonucleotide chain S2 is connected to at least one of a fluorescent marker, biotin, digoxigenin, and a nanoluminescent material.

3. A field effect transistor sensor for detecting novel coronavirus nucleic acid, characterized in that: include: A field effect transistor device and the Y-shaped DNA probe according to claim 1, wherein a semiconductor layer is provided on the field effect transistor device, and the Y-shaped DNA probe according to claim 1 is modified and fixed on the semiconductor layer.

4. A method for preparing a field effect transistor sensor for detecting novel coronavirus nucleic acid according to claim 3, characterized in that: obtaining a field effect transistor device; The field effect transistor device is placed in an ethanol solution of 3-aminopropyltriethoxysilane (APTES), incubated, and then heated to obtain an APTES-modified FET; the molar fraction of the ethanol solution of 3-aminopropyltriethoxysilane (APTES) is 5% to 8%; The APTES-modified FET is immersed in a 1-3 mg / mL sulfosuccinimidyl cyclohexane-1-carboxylate sodium salt (sulfo-SMCC) solution for incubation to obtain a sulfo-SMCC-functionalized FET; The sulfo-SMCC-functionalized FET was immersed in a solution of oligonucleotide chain S1 and incubated, and then immersed in a solution of oligonucleotide chain S2 and incubated after rinsing to obtain a field effect transistor sensor for detecting novel coronavirus nucleic acid.

5. The preparation method according to claim 4, characterized in that The field effect transistor device is placed in an ethanol solution of 3-aminopropyltriethoxysilane (APTES) and incubated for 50 to 60 minutes. The heating condition is 105° C. to 115° C. for 25 to 35 minutes.

6. The preparation method according to claim 4, characterized in that The APTES-modified FET is immersed in a cyclohexane-1-carboxylic acid sulfosuccinimidyl ester sodium salt (sulfo-SMCC) solution and incubated for 1.5 to 2 hours.

7. The preparation method according to claim 4, characterized in that The sulfo-SMCC functionalized FET is immersed in the solution of oligonucleotide chain S1 and incubated for 10 to 12 hours. After washing, it is immersed in the solution of oligonucleotide chain S2 and incubated for 0.5 to 1 hour.

8. Use of the Y-shaped DNA probe according to claim 1 or 2 or the field effect transistor sensor according to claim 3 in the preparation of a novel coronavirus nucleic acid detection product.

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