A method for detecting the pathogenicity of influenza A h1n1 virus based on pyrosequencing

Abstract

The invention relates to a method for detecting the pathogenicity of an influenza A (H1N1) virus based on pyrosequencing. The method comprises the steps of carrying out RT-PCR (Reverse Transcription-Polymerase Chain Reaction) amplification on a hemagglutinin (HA) gene of an H1N1 virus; carrying out pyrosequencing on a PCR amplification product to judge whether the cleavage site of the HA gene of the influenza A (H1N1) virus has a mutation, wherein the pyrosequencing is carried out in an SQA (Sequence Analysis) mode, and a nucleotide sampling sequence is shown as AGCT. The mutation of the cleavage site of the HA gene of the influenza A (H1N1) virus can be detected at rather high accuracy through comparing a detected result with the sequence of the cleavage site of the HA gene of a standard strain of the influenza A / H1N1 virus. The invention provides a simple and rapid experimental scheme for determining the virulence, pathogenicity and host range of the virus, the complex experiment steps related to complete genome sequencing are omitted, the variation direction of the virus can also be accurately mastered, the virus is conveniently monitored, an infection source is isolated, a transmission route is cut off, and the further development of an epidemic situation is stopped.

Description

technical field [0001] The invention relates to a method for detecting the pathogenicity of influenza A (H1N1) virus, in particular to a method for detecting the pathogenicity of influenza A (H1N1) virus based on pyrosequencing, and belongs to the technical field of virus detection. Background technique [0002] Influenza A (H1N1), which broke out in March 2009, was another major public health event after SARS and highly pathogenic avian influenza, which caused great panic to people all over the world. Influenza A virus is a single-strand negative-segmented RNA virus, which is prone to antigenic drift, antigenic conversion and genetic recombination, especially the mutation and recombination of the viral outer membrane protein hemagglutinin (HA), which can form various mutations Strains or recombinant strains lead to changes in the virulence and host range of the virus, thereby causing influenza epidemics of varying degrees. Whether hemagglutinin (HA) can be cut into HA1 and...

AI Technical Summary

Problems solved by technology

[0004] In order to strengthen the control of the rapid spread of H1N1 and avoid its outbreak, the development of rapid, reliable and high-throughput virus mutation detection technology is a research hotspot. The traditional automatic DNA sequencing technology based on the Sanger method is time-consuming in the detection of virus gene mutations. , laborious disadvantages

Pyrosequencing technology is a new generation of short-segment DNA sequence analysis technology, which can perform short DNA sequence analysis quickly, accurately and in real time, with high detection throughput, simple and convenient operation, and programmable. There is no report on the detection of the cleavage site of HA gene

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