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Marker nucleic acid probe, preparation method thereof, test strip and application of polypyrrole nanoparticles

A nucleic acid probe and pyrrole nanotechnology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as low sensitivity, and achieve the effects of improving detection sensitivity, high photostability, and high storage stability

Pending Publication Date: 2022-04-29
ANHUI SCI & TECH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the sensitivity of using colloidal gold as a labeling material is on the low side. According to literature, the minimum detection concentration of nucleic acid chromatography test strips based on colloidal gold is 500pM

Method used

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  • Marker nucleic acid probe, preparation method thereof, test strip and application of polypyrrole nanoparticles
  • Marker nucleic acid probe, preparation method thereof, test strip and application of polypyrrole nanoparticles
  • Marker nucleic acid probe, preparation method thereof, test strip and application of polypyrrole nanoparticles

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preparation example Construction

[0040] The present application also provides a method for preparing the above-mentioned marker nucleic acid probe, comprising the following steps:

[0041] S1, providing a dispersion of polypyrrole nanoparticles;

[0042] S2, sequentially adding deoxynucleotides, a first stabilizer and a detection probe to the dispersion to perform a coupling reaction, so that the detection probe is coupled to the polypyrrole nanoparticles.

[0043] In some embodiments, in step S1, the polypyrrole nanoparticles are dispersed in deionized water to obtain the dispersion, and the mass fraction of the polypyrrole nanoparticles in the dispersion is 1.5wt‰˜2.5wt‰.

[0044] In some embodiments, the preparation method of the polypyrrole nanoparticles is as follows: adding pyrrole monomers to the first solution containing the second stabilizer, then adding an oxidant to carry out oxidative polymerization, and finally adding a terminator to terminate the reaction, centrifuging Purifying the reacted sol...

Embodiment 1

[0071] Polypyrrole nanoparticle dispersion preparation: add water 100mL in 500mL Erlenmeyer flask, PVA (M w =31000) 8g, stirred at 900rpm for 20min at room temperature; added 500mg of pyrrole monomer, stirred at 900rpm for 20min at room temperature; quickly added 50mL (56mg / mL) of ferric chloride aqueous solution, stirred at 1500rpm for 6h at room temperature, added 20mL of methanol to terminate the reaction, Centrifuge at 15000rpm for 30min to remove the supernatant, wash the gained solid three times with hot water, and then disperse it in water for subsequent use. The mass fraction of polypyrrole nanoparticles in the dispersion is 2.5wt‰, and the average particle diameter of polypyrrole nanoparticles is 46nm, such as Figure 8 shown.

Embodiment 2

[0073] Polypyrrole nanoparticle dispersion preparation: add water 100mL in 500mL Erlenmeyer flask, PVA (M w =9000) 8g, stirred at 900rpm for 20min at room temperature; added 500mg of pyrrole monomer, stirred at 900rpm for 20min at room temperature; quickly added 50mL of ferric chloride aqueous solution (56mg / mL), stirred at 1500rpm for 6h at room temperature, and added 20mL of methanol to terminate the reaction. Centrifuge at 15000rpm for 30min to remove the supernatant, wash the gained solid three times with hot water, and then disperse it in water for subsequent use. The mass fraction of polypyrrole nanoparticles in the dispersion is 2.5wt‰, and the average particle diameter of polypyrrole nanoparticles is 89nm, such as Figure 9 shown.

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Abstract

The invention discloses a marker nucleic acid probe and a preparation method thereof, a test strip and application of polypyrrole nanoparticles. Wherein the marker nucleic acid probe comprises a marker and a detection probe loaded on the marker, and the marker is polypyrrole nanoparticles. According to the lateral chromatography test paper, the polypyrrole nanoparticles are creatively adopted as the marker, the polypyrrole nanoparticles have relatively high storage stability, relatively high conductivity and high light stability, and the lateral chromatography test paper based on the polypyrrole nanoparticles can be used for carrying out sensitive and quantitative visual detection on DNA or RNA in a short time; the minimum DNA concentration which can be detected by naked eyes is 1pM, so that the detection sensitivity of the lateral chromatography test paper is greatly improved.

Description

technical field [0001] The present application relates to the field of nucleic acid chromatography test paper, in particular to the use of marker nucleic acid probes and preparation methods thereof, test strips and polypyrrole nanoparticles. Background technique [0002] Detection of genes (DNA or RNA) is very important in gene therapy, clinical diagnosis and various biomedical research. The currently accepted method for genetic testing is the polymerase chain reaction (PCR). PCR is highly sensitive and accurate, but it is laboratory-based and requires relatively well-trained personnel. Additionally, false positive results often occur when small amounts of fragmented DNA are present as contaminants. Therefore, it is an urgent need to develop a simple, economical, highly sensitive and specific DNA rapid on-site detection technology. [0003] Lateral flow chromatography is a technique for detecting target substances in samples based on the hybridization reaction of nucleic ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6834C12Q1/6816C12Q1/6876
CPCC12Q1/6834C12Q1/6816C12Q1/6876C12Q2563/155C12Q2565/625C12Q2563/131Y02A50/30
Inventor 于庆才钱立生汪雁邱万伟李田田张静刘国东
Owner ANHUI SCI & TECH UNIV
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