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一株類禽型H1N1豬流感病毒的進化分析與分子特征

點擊數:6640  時間:2019-06-26 10:30:59  來源: 信息處      

2017年12月,上海市一養殖場飼養的豬出現咳嗽、呼吸困難、發熱及迅速轉歸等病症。将豬鼻拭子接種雞胚進行病毒分離,對血凝試驗陽性樣品在SPF雞胚上進一步純化、增殖,分離到1株豬流感病毒(A/swine/Shanghai/1205/2017)。采用RT-PCR對分離毒株進行全基因組擴增測序,利用DNAstar軟件,對測序基因片段進行整個閱讀框的核苷酸序列同源性比對分析,并用MEGA6繪制遺傳進化樹并分析氨基酸位點。結果顯示:分離毒株為H1N1亞型,其8個基因片段均屬于類禽型H1N1進化分支,沒有出現不同基因型流感病毒片段之間的重組;分離株HA蛋白的裂解位點序列為PSIQSR↓G,具有典型低緻病性流感病毒的分子特征。本毒株的分離鑒定為分析我國大陸地區的豬流感流行狀況和分子特征提供了參考。



Phylogenetic Analysis on a Strain of Avian-like H1N1 SubtypeSwine Influenza Virus and Determination of Its Molecular Characteristics

In December 2017,pigs in afarm of Shanghai City appeared a series of symptoms,includingcough,dyspnea and fever,and thediseased pigs recovered rapidly. Then the nasal swab samples collected frompigs were inoculated into SPF chicken embryos for virus isolation,after further purifying and proliferating the positive samplesdetected by haemagglutination assay(HA),one strain of swine influenza virus(A/swine/Shanghai/1205/2017)was isolated. The whole genome of the isolate was amplified andsequenced by RT-PCR,the nucleotide sequence homology ofthe whole reading frame was analyzed by the DNAstar software,and then the genetic evolution tree was designed using MEGA 6 toanalyze the amino acid sites. The results showed that the isolated strain wasidentified as H1N1 subtype influenza virus,and itseight gene fragments were classified into evolutionary branch of avian-likeH1N1,with no any recombination among influenza virusfragments of various genotypes. The cleavage site sequence of its HA proteinwas PSIQSR↓G,indicating thetypical molecular characteristics of low pathogenicity influenza virus.Therefore,the isolation and identification of H1N1strain would provide some reference for analyzing the epidemic situation andmolecular characteristics of swine influenza in Mainland China.

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