Indium tin oxide field effect transistor biosensor based on DNA nano quadruple and application of indium tin oxide field effect transistor biosensor

An indium tin oxide field, biosensor technology, applied in the directions of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problems of high false positive rate, long detection time, and complicated detection procedures in short-sequence detection. Improve nucleic acid hybridization efficiency, high amplification efficiency and strong specificity

Pending Publication Date: 2022-03-22
FUZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Among them, the RT-LAMP detection procedure is complex, the detection time is long (it takes several hours, and the detection results need to be checked), and false negative results often occur; the NEAR reaction mechanism is complex, and the output is easily limited by the template, and it is not suitable for the detection of short sequences. high false positive rate
[0004] Conventional field-effect transistor (FET) biosensors immobilize probes on the gate surface and use the principle of complementary base pairing to capture target nucleic acids, which makes vertical nucleic acid signals beyond the Debye length undetectable

Method used

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  • Indium tin oxide field effect transistor biosensor based on DNA nano quadruple and application of indium tin oxide field effect transistor biosensor
  • Indium tin oxide field effect transistor biosensor based on DNA nano quadruple and application of indium tin oxide field effect transistor biosensor
  • Indium tin oxide field effect transistor biosensor based on DNA nano quadruple and application of indium tin oxide field effect transistor biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Sensitivity Experiment of Indium Tin Oxide Field Effect Transistor Biosensor Based on DNA Nano-quadruplex

[0044] (1) Indium tin oxide field effect transistor (ITO FET) was treated with oxygen plasma, the ratio of argon to oxygen was 4:1, the power was 15 W, and the treatment was 2 min.

[0045] (2) Add the APTES solution to the ITO FET treated with oxygen plasma; the concentration of APTES is 5wt%, the solvent is a mixture of absolute ethanol and acetic acid, the content of acetic acid is 10vol%, and the pH value is 5. React at room temperature for 6 hours; after the reaction, clean the ITO FET with absolute ethanol and deionized water; blow dry with nitrogen, and bake at 110°C for 30 minutes.

[0046] (3) Prepare 25 μmol / L hairpin structure DNA solutions H1, H2, H3, H4 and streptavidin (SA) solution with 1×PBS buffer solution with pH=7.4, and refrigerate at 4°C for later use.

[0047] (4) Streptavidin (SA) solution was mixed with hairpin structure DNA H3 s...

Embodiment 2

[0053] Example 2 Indium Tin Oxide Field Effect Transistor Biosensor Based on DNA Nano-quadruplex

[0054] (1) Indium tin oxide field effect transistor (ITO FET) was treated with oxygen plasma, the ratio of argon to oxygen was 2:1, the power was 10 W, and the treatment was 10 min.

[0055] (2) Add the APTES solution to the ITO FET treated with oxygen plasma; the concentration of APTES is 5wt%, the solvent is a mixture of absolute ethanol and acetic acid, the content of acetic acid is 1vol%, and the pH value is 6. React at room temperature for 4 hours; after the reaction, clean the ITO FET with absolute ethanol and deionized water; blow dry with nitrogen, and bake at 120°C for 30 minutes.

[0056] (3) Prepare 20 μmol / L DNA solutions H1, H2, H3, H4, and streptavidin (SA) solution with 1×PBS buffer solution with pH=7.4, and refrigerate at 4°C for later use.

[0057] (4) Mix 20 μmol / L streptavidin (SA) solution with biotinylated DNA probe (H3 / H4) at a ratio of 1:4, and react for 1...

Embodiment 3

[0061] Example 3 Indium Tin Oxide Field Effect Transistor Biosensor Based on DNA Nano-quadruplex

[0062] (1) Indium tin oxide field effect transistor (ITO FET) is treated with oxygen plasma, the ratio of argon to oxygen is 5:1, the power is 50W, and the treatment is 6 minutes;

[0063] (2) Add the APTES solution to the oxygen plasma-treated ITO FET; the concentration of APTES is 2wt%, the solvent is a mixture of absolute ethanol and acetic acid, the content of acetic acid is 5vol%, and the pH value is 6. React at room temperature for 6 hours; after the reaction, clean the ITO FET with absolute ethanol and deionized water; blow dry with nitrogen, and bake at 115°C for 45 minutes;

[0064] (3) Prepare 50 μmol / L DNA solutions H1, H2, H3, H4, and streptavidin (SA) solution with 1×PBS buffer solution with pH=7.4, and refrigerate at 4°C for later use;

[0065] (4) Streptavidin (SA) solution and biotinylated DNA probe (H3 / H4) were mixed at a ratio of 1:4, and reacted for 15 minutes...

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Abstract

The invention discloses an indium tin oxide field effect transistor biosensor based on a DNA nano quadruple and application of the indium tin oxide field effect transistor biosensor. The biosensor comprises a hairpin structure capture probe DNA H1, a hairpin structure replacement probe DNA H2, a hairpin structure DNA H3, a hairpin structure DNA H4 and an indium tin oxide field effect transistor. The biosensor is combined with a nucleic acid hybridization product cHCR-SANTs of a biotin-streptavidin novel biological reaction amplification system, the cHCR-SANTs has a stable spatial network structure, and the cHCR-SANTs can continuously extend towards the periphery and are not perpendicular to the surface of a channel, so that the nucleic acid hybridization efficiency within the length of Debye is effectively improved; therefore, the charge change on the surface of the channel is greatly changed, and ultralow-concentration target nucleic acid detection is realized.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to an indium tin oxide field-effect transistor biosensor based on a DNA nanometer quadruplet and an application thereof. Background technique [0002] Testing for COVID-19 is crucial. Clinical testing is mainly for SARS-CoV-2 antibodies and nucleic acids. SARS-CoV-2 antibody detection is simple and fast, but the antibody has a certain window period and can only be used as a supplementary method for nucleic acid detection. As the "gold standard" for diagnosing pathogenic infection, nucleic acid detection has the characteristics of early diagnosis, high sensitivity and specificity, and is suitable for early screening of asymptomatic infections and management and control of patients' diseases. The nucleic acid detection methods of the novel coronavirus currently on the market include whole genome sequencing (NGS), real-time fluorescent reverse transcriptase c...

Claims

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Application Information

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IPC IPC(8): G01N27/414C12Q1/6825C12Q1/682C12Q1/70C12R1/93
CPCG01N27/4145C12Q1/6825C12Q1/682C12Q1/701C12Q2525/301C12Q2563/131C12Q2537/1373Y02A50/30
Inventor 李调阳林美雅林本慧王鸣巍
Owner FUZHOU UNIVERSITY
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