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
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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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