Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

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

A pyrosequencing, influenza virus technology, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of laborious, time-consuming detection of virus gene mutations, etc., to achieve simple operation, prevent the further expansion of the epidemic, and quickly The effect of the experimental protocol

Inactive Publication Date: 2016-05-18
山东国际旅行卫生保健中心
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for detecting the pathogenicity of influenza A h1n1 virus based on pyrosequencing
  • A method for detecting the pathogenicity of influenza A h1n1 virus based on pyrosequencing
  • A method for detecting the pathogenicity of influenza A h1n1 virus based on pyrosequencing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040] The content of the present invention will be further described in detail below by way of examples.

[0041] 1. RNA extraction

[0042] Use freshly propagated virus-infected MDCK passage cells as templates for RNA extraction, and use the HighPureViralRNAKit produced by Roche to extract viral RNA. The specific operation is carried out according to the kit instructions. The extracted RNA is immediately used for RT-PCR amplification or stored at -80°C. .

[0043] 2. RT-PCR reaction to amplify the HA1 gene fragment

[0044] The first round of amplification was carried out with the PCR primers having the sequence indicated by H1F and the PCR primers having the sequence indicated by H1R, and the extracted viral RNA was used as a template. In the 50μL reaction system, the optimal concentration of the first primer is 50nmol / L. The RT-PCR reaction conditions were: reverse transcription at 50°C for 30min; pre-denaturation at 94°C for 2min; 45 cycles of 94°C for 15s, 55°C for 30...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

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...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6869C12Q1/70C12Q1/701C12Q2600/142C12Q2600/156C12Q2531/113C12Q2565/301
Inventor 陈晓光程琳郭宁张瑾姜华金燕梁洁张娟
Owner 山东国际旅行卫生保健中心
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com