Kit for detecting H5N2 avian influenza virus

A bird flu virus and kit technology, applied in the field of biological detection, can solve the problems of lack of simple, fast and easy-to-use detection methods, large losses in breeding industry, etc., and achieve high sensitivity, low cost and simple operation

Inactive Publication Date: 2013-01-02
天津朝海科技有限公司
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  • Abstract
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AI Technical Summary

Problems solved by technology

Current data show that H5N2 avian influenza, regardless of its high pathogenicity or low pathogenicity, is internationally recognized as a poultry disease and will not infect humans, but it has caused huge losses to the breeding industry
[0007] At present, there is no simple, rapid, easy-to-use and low-cost detection method for H5N2 avian influenza virus

Method used

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  • Kit for detecting H5N2 avian influenza virus

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Embodiment Construction

[0023] The kit of the present invention includes 2 tubes of solutions, one tube is 50 μl of a nano-gold particle solution labeled with a molecular probe that can recognize the specific sequence of the H5N2 type avian influenza virus, and the other tube is 250 μl of a solution that can amplify the DNA in the sample to be detected. H5N2 avian influenza virus DNA or a loop-mediated isothermal nucleic acid amplification system that amplifies H5N2 avian influenza virus RNA in a sample to be detected after a reverse transcription process.

[0024] The gold nanoparticle solution contains gold nanoparticles with a diameter of 20 nm at a concentration of 1.2 nM, and the surface of the gold nanoparticles is marked with thiol-modified DNA probes.

[0025] The components of the loop-mediated isothermal nucleic acid amplification system are as follows: 0.2 μM F3 and B3 primers, 1.6 μM FIP and BIP primers, 0.4 M betaine (Sigma–Aldrich, St. Louis, MO), 10 mM MgSO4, 1.4 mM dNTPs, 1× ThermoPol...

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Abstract

The invention relates to a kit adopting gold nanoparticle to detect DNA formed by amplifying H5N2 avian influenza virus DNA through the loop-mediated isothermal nucleic acid amplification technology, and belongs to the technical field of biological detection. The kit combines the loop-mediated isothermal nucleic acid amplification technology and the bio-nanotechnology, improves the sensitivity of biomolecule detection, is simple, rapid and accurate to operate, does not require special instrument and equipment, and can be widely applied to the detection of high-sensitivity H5N2 avian influenza virus in fields of family diagnosis, clinical diagnosis, communicable disease control, environment monitoring, inspection and quarantine, biotechnology and the like. The kit comprises two parts; the first part is gold nanoparticle, on which a molecular probe capable of identifying H5N2 avian influenza virus specific sequences is marked; and the second part is a loop-mediated isothermal nucleic acid amplification system capable of amplifying H5N2 avian influenza virus DNA in samples to be detected or capable of amplifying H5N2 avian influenza virus RNA after a reverse transcription process.

Description

technical field [0001] The invention relates to a kit for detecting H5N2 avian influenza virus DNA or RNA amplified by a loop-mediated isothermal nucleic acid amplification technology by using nano gold particles, and belongs to the technical field of biological detection. Background technique [0002] The high sensitivity, high specificity, simple and rapid detection of genes (DNA) and their transcription products (RNA) is very useful for the early diagnosis and treatment of diseases, as well as in the fields of health and epidemic prevention, environmental monitoring, inspection and quarantine, etc. Significance. In the past, more accurate and sensitive detection methods in this regard include fluorescent labeling, radioactive labeling and other methods. These methods require complex operations and special equipment, and their application range is very narrow. In 1993, Dr. Kary B. Mullis of PE Company invented the polymerase chain reaction (Polymerase Chain Reaction), ref...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 不公告发明人
Owner 天津朝海科技有限公司
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