2014年长沙市活禽市场污水中H5N1亚型禽流感病毒HA、NA及NS基因进化分析

发布时间：2018-07-10 08:20

本文选题：活禽市场 + 环境　；参考：《中国人兽共患病学报》2017年01期

【摘要】：目的分析2014年长沙市活禽市场污水中H5N1亚型禽流感病毒(Avian influenza virus,AIV)血凝素(Hemagglutinin,HA)、神经氨酸酶(Neuraminidase,NA)及非结构蛋白(Non-structural,NS)基因进化特征。方法从2014年长沙市活禽市场采集501份环境标本,利用Real-time PCR方法进行A型、H5、H7和H9亚型流感病毒核酸检测,并对单一H5阳性标本进行核苷酸测序,病毒HA、NA和NS基因测序结果进行在线BLAST分析,利用Mega5和Bioedit软件构建核苷酸进化树和氨基酸(Amino acids,aa)比对。结果从501份环境标本中检出A型H5亚型病毒阳性标本177份(检出率35.33%),其中8份标本经核苷酸测序鉴定为H5N1亚型病毒,进化分析表明大部分H5N1病毒HA基因位于2.3.2分支内,聚集形成一个新亚分支,NA及NS基因位于2.3.2.1b亚分支。HA蛋白受体结合位点aa为QSG,表现为禽流感病毒受体特征;NA蛋白未出现H275Y及N295Saa替换,对神经氨酸酶抑制剂敏感;HA、NA及NS蛋白关键分子位点表现为高致病性的分子特征。结论 2014年长沙市活禽市场污水中H5N1亚型禽流感病毒HA、NA和NS基因表现为高致病性的分子特征,但不具有对人易感的受体特征,需要进一步监测。
[Abstract]:Objective to analyze the evolutionary characteristics of hemagglutinin (HA), neuraminidase (na) and non-structural protein (NNS) genes of H5N1 subtype avian influenza virus (Avian influenza virusus AIV) in the wastewater from live poultry market in Changsha City in 2014. Methods 501 environmental samples were collected from live poultry market in Changsha in 2014. Real-time PCR was used to detect the nucleic acid of influenza virus type A H5H7 and H9 subtype, and the nucleotide sequence of single H5 positive sample was sequenced. The results of Hana and NS gene sequencing were analyzed by blast on line. The nucleotide evolutionary tree and amino acid (Amino acidsof) were constructed by Mega5 and Bioedit software. Results 177 (35.33%) positive samples of H5 subtype A virus were detected from 501 environmental samples, 8 of them were identified as H5N1 subtype virus by nucleotide sequencing. Evolution analysis showed that most of the HA gene of H5N1 virus was located in the branch of 2.3.2. A new subbranch, na and NS gene, located at 2.3.2.1b subbranch. Ha protein receptor binding site AA was QSG. it was shown that there was no H275Y and N295Saa substitution in the Avian Influenza virus receptor characteristic. The key molecular sites of Hana and NS protein were highly pathogenicity to neuraminidase inhibitors. Conclusion the Hana and NS genes of H5N1 subtype avian influenza virus in Changsha live poultry market wastewater in 2014 are highly pathogenic, but they do not possess the receptor characteristics of susceptibility to human, and need to be monitored further.
【作者单位】：长沙市疾病预防控制中心;
【基金】：湖南省卫生计生委2015年度科研计划项目(B2015-153)
【分类号】：R511.7

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