猪流感病毒广东株分离鉴定及遗传进化分析

doi:10.16431/j.cnki.1671-7236.2020.08.032

Abstract

Abstract: In this study,37 swabs were collected from a pig farm with suspected swine influenza in Guangdong province,inoculated into 9-day-old SPF chicken embryos and collected alluvial fluid,and a strain of swine influenza virus was isolated by hemagglutination (HA) test and hemagglutination inhibition (HI) test and RT-PCR identification.Eight gene fragments were amplified for sequencing and sequence analysis,and then compared with the reference strains in GenBank to construct the evolutionary tree.The results showed that the virus strain was H1N1 swine influenza virus.The isolate was designated as A/swine/Guangdong/2/2018 (H1N1).The complete genes were sequenced and the genetic analysis showed that the nucleotide sequences of the 8 segments of the isolate were similar to the corresponding sequences of A/swine/Guangdong/L3/2009(H1N1) by more than 99%,and were in the same branch as the classical swine influenza virus of the H1N1 subtype.The cleavage site of HA gene of the isolate was PSIQS↓GL,which was consistent with the molecular characteristics of low pathogenic influenza virus.HA gene receptor sites were 190D,225G and 226Q,which indicated that the strain could bind to both SA receptor of SAα-2,6-Gal human influenza virus and SA receptor of SAα-2,3-Gal avian influenza virus.There were six potential glycosylation sites at 28,40,104,304,498 and 557 amino acids.There were six potential glycosylation sites in amino acids of 50,58,63,68,98,146 and 235 of NA protein.The active sites of amino acid sequence of NA protein were 119E,199D,223I,275H,293R and 295N.There was no mutation in amino acid analysis site,indicating that the isolate was highly sensitive to neuraminidase inhibitors,but in M2 protein,31 amino acids changed from sensitive (S) to resistant (N),which might be resistant to amantadine drugs.The evolution analysis of swine influenza virus would provide important information for the prevalence and variation of swine influenza in Guangdong province.

Key words: swine influenza; H1N1 subtype; isolation and identification; genetic evolution

CLC Number:

S855.3

HUANG Liangzong, YAN Guangzhi, DENG Rusen, CHEN Shengnan, ZHANG Hailong, GU Wanjun. Isolation,Identification and Genetic Evolution Analysis of Swine Influenza Virus from Guangdong Province[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2625-2633.

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Isolation,Identification and Genetic Evolution Analysis of Swine Influenza Virus from Guangdong Province

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