Swine flu virus identification primer and application thereof

A swine influenza virus, influenza virus technology, applied in the direction of microorganisms, microorganism-based methods, microorganism determination/inspection, etc., to achieve the rapid effect of classical virus isolation and identification and serological typing test

Inactive Publication Date: 2014-03-12
山东省动物疫病预防与控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no diagnostic method established for different lineages and subtypes of swine influenza virus at present

Method used

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  • Swine flu virus identification primer and application thereof
  • Swine flu virus identification primer and application thereof
  • Swine flu virus identification primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1, the design of primer

[0075] 1. Design of primers

[0076] The published HA genes registered in Genbank belong to classical H1 subtype swine influenza virus strains (Genbank accession numbers GU086009, EU502885, GU086041, GU086033, GU086081, EF556199, GQ117044, ADT79242), European avian H1 subtype swine influenza virus ( Genbank accession numbers FJ536762, FJ536810), human-like H3 subtype swine influenza virus (Genbank accession numbers GU086089, EU655695, GU086097, EU655692), H9 subtype swine influenza virus (Genbank accession numbers DQ981586, DQ981546), H5 subtype influenza virus ( Genbank accession numbers DQ100557, DQ997392, DQ997076, AY747609, DQ432037, DQ997262, AY646424) highly conserved sequence, design 5 pairs of specific primers (SH3-420F, SH3-614R; SEAH1-218F, SEA-H1-415R; SCH1-153F , SCH1-534R; SH9-643F, SH9-1160R; SH5-379F, SH5-1063R). The expected amplified fragment sizes are H3A: 195bp; SEA-H1A: 278bp; SC-H1A: 382bp; S-H9A: 518bp; S-H5A:...

Embodiment 2

[0125] Embodiment 2, the application of primer

[0126] 1. Treatment of diseased materials to be tested: resuspend the nasal test papers of suspected diseased pigs (derived from scattered farmers such as Guangdong Province) numbered 1-7 with sterilized water, and extract RNA. The method is the same as that of 2 in Example 1. In step (1), the RNA of pigs numbered 1-7 is obtained.

[0127] 2. Multiplex PCR detection

[0128] Reverse transcribe the above-mentioned pig RNA numbered 1-7 to obtain cDNA, the method is the same as step (2) of 2 of Example 1, and obtain the pig cDNA numbered 1-7 respectively.

[0129] Using the cDNA of the above-mentioned pigs numbered 1-7 as templates and 5 pairs of primers as primers, PCR amplification was carried out, and the amplification system and amplification conditions were the same as step (3) in 2 of Example 1.

[0130] If the amplified product is a 684bp fragment, the pig to be tested is or is a candidate for H5 subtype influenza virus in...

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Abstract

The invention discloses a swine flu virus identification primer and an application thereof. The swine flu virus identification primer provided by the invention consists of primer groups A to E, wherein the primer group A consists of a primer 1 and a primer 2, the primer group B consists of a primer 3 and a primer 4, the primer group C consists of a primer 5 and a primer 6, the primer group D consists of a primer 7 and a primer 8, the primer group E consists of a primer 9 and a primer 10, nucleotide sequences of the primer 1 to the primer 10 are respectively the sequence 1 to the sequence 10 in a sequence table. Experiments prove that the primer groups provided by the invention are characterized in that the subtype and the lineage of the swine flu virus is identified in one step through extracting sample ribonucleic acid (RNA), synthesizing complementary deoxyribonucleic acid (cDNA) via reverse transcriptase and simultaneously amplifying the genes with the possibly existence of H1, H3, H9 and H5 in one step. Compared with the typical virus separation identification and serology grouping tests, the primer has the advantage that the speed and the accuracy are high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer for identifying swine influenza virus and its application. Background technique [0002] Swine influenza (Swine Influenza, SI) is an acute contagious respiratory disease of pigs caused by type A influenza virus, which is characterized by sudden onset, cough, fever, dyspnea, exhaustion, and can also lead to pig breeding such as abortion in pregnant mothers. obstacle. Although the mortality rate of SI is not high, it affects the development of pigs and delays the growth and slaughter time of pigs; SI is also an immunosuppressive disease that can destroy the immune system of pigs and cause immunosuppression, leading to concurrent or secondary diseases of other diseases Infection, aggravating the disease, causing significant economic losses to the pig industry. Since the classic H1N1 subtype SI was clinically discovered in 1918, new subtypes and genotype strains have continuou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12R1/93
Inventor 刘金华毕玉海陈静田夫林张进林李云岗兰邹然
Owner 山东省动物疫病预防与控制中心
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