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Test strip biological sensor based on carbon nano tubes and triple helix molecular switch and preparation method and application

A biosensor, carbon nanotube technology, applied in biochemical equipment and methods, biological testing, and microbial determination/inspection, etc., can solve the problems of small design flexibility, poor specificity, low sensitivity, etc. Low production cost and strong resistance to denaturation

Inactive Publication Date: 2019-02-01
ANHUI SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the design flexibility of the current lateral flow nucleic acid (LFNA) test strip target sequence is small, the specificity is poor, and the sensitivity is low.

Method used

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  • Test strip biological sensor based on carbon nano tubes and triple helix molecular switch and preparation method and application
  • Test strip biological sensor based on carbon nano tubes and triple helix molecular switch and preparation method and application
  • Test strip biological sensor based on carbon nano tubes and triple helix molecular switch and preparation method and application

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preparation example Construction

[0030] The present invention also provides a preparation method of the test strip biosensor, comprising the following steps: 1) ultrasonically treating carbon nanotubes under acidic conditions to obtain carboxylated carbon nanotubes; 2) making the carboxylated carbon nanotubes Dehydration and condensation of the tube and the first amino DNA to obtain carbon nanotubes bound to the first amino DNA; 3) mixing the carbon nanotubes bound to the first amino DNA and single-stranded DNA P0 in an incubation solution to obtain triple helix binding Carbon nanotubes with DNA molecular structure; 4) Mix and spray the carbon nanotubes with the triple-helix DNA molecular structure described in step 3) and the carbon nanotubes with the second amino DNA on the binding pad; 5) Apply the first DNA-biological The first DNA-biotin-streptavidin complex and the second DNA-biotin-streptavidin complex were obtained by mixing and incubating with streptavidin respectively; 6) The first DNA-biotin-strept...

Embodiment 1

[0042] Use mixed concentrated acid (concentrated sulfuric acid: concentrated nitric acid = 3:1) to sonicate multi-walled carbon nanotubes (MWCNTs, hereinafter referred to as CNTs) with a tube length of 10 μm and a diameter of 30-50 nm for 12 hours, and use them after centrifugation (14500 rpm, 20 min). The PBS solution was washed until the CNTs solution was neutral, and the solution concentration was 1 mg / ml.

[0043] Take 500 μL of the above CNTs solution, add 500 μL of secondary water to form a 1 mL system, mix well, centrifuge at 14,000 rpm for 25 min to remove the supernatant, dissolve in 1 mL of MES buffer solution (pH=4.7) containing 9.6 mg of EDC and 5.4 mg of NHS, shake at room temperature bed for 1 h, washed with PBS buffer solution for 3 times, dissolved in 1 mL of PBS (pH=7.4), added 0.1 OD of the first amino DNA (5'-NH2-C6-AGAGGGAAA-3', Shanghai Sangong), shaker After overnight reaction, centrifugation (14500rpm, 20min) and washing with PBST three times, the precip...

Embodiment 2

[0054] Detection of carcinoembryonic protein CEA using a dipstick biosensor based on carbon nanotubes and a triple-helix molecular switch.

[0055] The designed partial sequence of single-stranded DNA P0 is the aptamer of CEA. Except for the following parts, other parts of the test strip are the same as in Example 1: the nucleotide sequence of the single-stranded DNA P0 is 5'-TCCCTTTCCCATAGGGAAGTGGGGGATTTCCCT-3' (SEQ ID No.6).

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Abstract

The invention provides a test strip biological sensor based on carbon nano tubes and a triple helix molecular switch and a preparation method and application, and belongs to the technical field of nucleic acid biological sensors. The test strip biological sensor based on the carbon nano tubes and the triple helix molecular switch comprises a base plate, a sample pad, a combined pad, a detection pad and an adsorption pad, wherein the sample pad, the combined pad, the detection pad and the adsorption pad are sequentially connected to the base plate in a building mode; the combined pad is coatedwith a carbon nano tube by combining a triple helix DNA molecular structure and a carbon nano tube by combining a second amidogen DNA; a detection line and a quality control line are arranged on the detection pad; the detection line is coated with a first DNA-biotin-streptavidin compound, the quality control line is coated with a second DNA-biotin-streptavidin compound, and complementary pairing of a single stranded DNA P0 sequence with a nucleotide sequence of nucleic acid to be detected is achieved. The test strip biological sensor has high sensitivity, good specificity and universality.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid biosensors, and in particular relates to a test strip biosensor based on carbon nanometer and triple helix molecular switch and its preparation method and application. Background technique [0002] Nucleic acids are important biological macromolecules that encode and regulate the expression of genetic information in all living things. However, some small changes in nucleic acid sequences can lead to biomedically and biologically important changes. Nucleic acid testing is essential for the diagnosis and treatment of genetic diseases, detection of infectious agents, drug discovery or warning of biological warfare agents. Conventional methods such as Northern, Southern and Western blots, agarose and polyacrylamide gel electrophoresis do not provide enough specific information and are very laborious and time consuming, requiring up to 48 hours of meticulous laboratory work. New methods and tech...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C12Q1/6804
CPCC12Q1/68G01N33/6893
Inventor 刘国东黄炎钱立生张学记
Owner ANHUI SCI & TECH UNIV
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