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A method for rapid identification of n1 and n2 subtype influenza viruses

An influenza virus, rapid technology, applied in the field of identifying different subtypes of influenza virus, can solve the problem of lack of NA subtype detection technology

Inactive Publication Date: 2018-12-25
POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, most of the methods for detecting AIV are aimed at the detection of its HA subtypes, such as H3, H4, H5, H7, H9 and other subtypes. There are few reports on the detection technology for NA subtypes, especially for the identification of multiple NA subtypes. diagnosis

Method used

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  • A method for rapid identification of n1 and n2 subtype influenza viruses
  • A method for rapid identification of n1 and n2 subtype influenza viruses
  • A method for rapid identification of n1 and n2 subtype influenza viruses

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Embodiment 1

[0041] Materials and Methods

[0042] 1. Materials

[0043] 1. Virus template: N1 is the total RNA obtained by mixing and extracting equal amounts of two identified strains of H5N1, and N2 is the total RNA obtained by mixing and extracting equal amounts of samples from one identified strain of H5N2 and one strain of H9N2.

[0044] 2. Main reagents: Small plasmid extraction kit and One-step Real-time RT-PCR kit were purchased from TaKaRa Company.

[0045] 3. Design and synthesis of primers and probes: select the highly conserved regions of the above strains N1 and N2 genes, and use PrimerExpress to design primers and probes. The fluorescent reporter group labeled at the 5' end of the N1 probe is FAM, and the quencher group labeled at the 3' end is BQ1; the fluorescent reporter group labeled at the 5' end of the N2 probe is VIC, and the quencher group labeled at the 3' end is The group was BQ2, and the primers and probes were synthesized by Life Company. The primers were purif...

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Abstract

The invention discloses a method for quickly detecting N1 and N2 subtype influenza viruses. According to the invention, a one-step fluorescence quantitative RT-PCR detection method is adopted, Buffer, dNTP and Enhance solution are premixed to form 2X one-step buffer, and multiple enzymes are mixed according to a ratio. Preparation of reaction liquids of the operation is simple and convenient, the operation time is shortened, and meanwhile operation pollution is avoided. The method disclosed by the invention is simple and convenient to operate, good in sensibility, convenient and quick, is suitable for developing a fluorescence quantitative RT-PCR kit for identifying and detecting N1 and N2 subtypes, is used for N1 and N2 subtype clinical differential diagnosis and epidemiological survey, and is of great significance for prevention and control of influenza in China.

Description

technical field [0001] The invention relates to a method for identifying different subtypes of influenza virus, in particular to a method for quickly identifying N1 and N2 subtype influenza viruses by using a multiplex fluorescent quantitative PCR method. Background technique [0002] Influenza Virus belongs to Orthomyxoviridae family and is an RNA virus. According to the differences of viral membrane protein (M) and nucleoprotein (NP), influenza viruses are divided into three large groups: A (A), B (B), and C (C). Among them, influenza A has the longest epidemic time, the greatest harm, and the widest range of epidemic hosts. According to the structure of hemagglutinin (HA) and neuraminidase (NA) on the surface of the virus and their genetic characteristics, they can be divided into different subtypes. At present, there are 16 subtypes of hemagglutinin (HA) found in influenza A virus, represented by H1, H2...H16 respectively; 9 subtypes of neuraminidase, represented by NI...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6851
CPCC12Q1/686C12Q1/70C12Q2537/143C12Q2563/107
Inventor 王友令袁小远宋敏训艾武徐怀英亓丽红张玉霞
Owner POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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