2015-2017年贵州省威宁13株H9N2亚型禽流感病毒HA基因序列分析
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摘要
目的了解2015-2017年贵州省威宁H9N2亚型禽流感病毒的分子特征,为禽流感病毒的研究与防控提供依据。方法对选取的13株病毒进行核酸的提取、血凝素基因(hemagglutinin,HA)的扩增和测序,运用生物信息学软件对H9N2禽流感病毒(AIV) HA基因的同源性、遗传进化、受体结合关键位点、致病相关位点和糖基化位点变异情况进行分析。结果 13株H9N2AIV的HA基因核苷酸和氨基酸同源性分别为96. 1%～99. 9%和95. 7%～100%,均属于DK/HK/Y280/97分支。HA基因与致病性高低相关的裂解位点区域2株为PSRSNR↓GLF,11株为PSRSSR↓GLF,均属于低致病性毒株。与传播感染相关的受体结合位点均发生E198T和Q234L的突变,具有人样受体结合特征。13株毒株HA均含7个与毒力相关的糖基化位点,218位点均存在一个糖基化位点缺失,313位点均存在一个糖基化位点的增加。结论贵州省威宁县H9N2AIVs属于DK/HK/Y280/97系,均为低致病性禽流感病毒,存在人易感位点,病毒一直处于不断的变异之中,故应加强其监测与防控。
Objective To understand the molecular characteristics of the hemagglutinin gene( HA) of H9N2 subtype avian influenza virus( AIV) in Weining from 2015 to 2017,and to provide scientific evidence for the prevention and control of AIV. Methods RNAs were extracted and the HA genes from the 13 samples with H9N2 positive screened by Real-Time PCR of live poultry market( LPM) environment were sequenced. Then the homology,genetic evolution,pivotal sites related to receptor binding regions,pathogenicity and potential glycosylation of H9N2 subtype AIV were analyzed by a series of bioinformatic softwares. Results 96. 1% ～ 99. 9% and 95. 7% ～ 100% strains were similar among 13 strains in nucleotide and amino acid of the HA gene,respectively. All strains belonged to Eurasian lineage,DK/HK/Y280/97 sublineage. The cleavage sites motif of2 strains was PSRSNR↓GLF and that of 9 strains was PSRSSR↓GLF,both belonging to low pathogenic strains. Mutations in E198 T and Q234 L at the receptor binding sites in the HA were found in All strains indicated the potential of binding with the human-like receptor. All 13 strains had 7 glycosylation sites,a site deletion at amino acid site 218,and addition at313. Conclusion All H9N2 strains belong to DK/HK/Y280/97 sublineage,and they are low pathogenic avian influenza viruses in Weining,but mutations of receptor binding site may enhance the susceptibility and pathogenicity to human beings,and H9N2 virus is continual changing. Hence,enhancing the surveillance and control of H9N2 virus is necessary.
Objective To understand the molecular characteristics of the hemagglutinin gene( HA) of H9N2 subtype avian influenza virus( AIV) in Weining from 2015 to 2017,and to provide scientific evidence for the prevention and control of AIV. Methods RNAs were extracted and the HA genes from the 13 samples with H9N2 positive screened by Real-Time PCR of live poultry market( LPM) environment were sequenced. Then the homology,genetic evolution,pivotal sites related to receptor binding regions,pathogenicity and potential glycosylation of H9N2 subtype AIV were analyzed by a series of bioinformatic softwares. Results 96. 1% ～ 99. 9% and 95. 7% ～ 100% strains were similar among 13 strains in nucleotide and amino acid of the HA gene,respectively. All strains belonged to Eurasian lineage,DK/HK/Y280/97 sublineage. The cleavage sites motif of2 strains was PSRSNR↓GLF and that of 9 strains was PSRSSR↓GLF,both belonging to low pathogenic strains. Mutations in E198 T and Q234 L at the receptor binding sites in the HA were found in All strains indicated the potential of binding with the human-like receptor. All 13 strains had 7 glycosylation sites,a site deletion at amino acid site 218,and addition at313. Conclusion All H9N2 strains belong to DK/HK/Y280/97 sublineage,and they are low pathogenic avian influenza viruses in Weining,but mutations of receptor binding site may enhance the susceptibility and pathogenicity to human beings,and H9N2 virus is continual changing. Hence,enhancing the surveillance and control of H9N2 virus is necessary.
引文
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[12]叶贺佳,梁昭平,彭特,等. 2012-2015年H9N2亚型AIV分离株HA基因的克隆及序列分析[J].中国畜牧兽医,2016,43(5):1148-1155.
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[14]卢受昇,高慧敏,邓国东,等.广东地区近2年禽流感H_9亚型HA基因的序列分析[J].广东畜牧兽医科技,2014,39(6):29-32,41.
[2]周虹妤,裴磊,毛爱民,等. H9N2病毒研究进展[J].中国畜牧兽医文摘,2016,32(6):152.
[3] Sun YP,Pu J,Jiang ZL,et al. Genotypic evolution and antigenic drift of H9N2 influenza viruses in China from 1994 to 2008[J].Veterinary Microbiology,2010,146(3/4):215-225.
[4]丛彦龙,钟颖,孙艺学,等. H9N2亚型禽流感病毒流行病学及其疫苗的研究进展[J].中国兽医学报,2017,37(2):386-392.
[5]刘晓蕾,袁洁,欧新华,等.长沙市首例人感染H9N2禽流感病毒的分离及基因特征分析[J].现代预防医学,2016,43(17):3225-3231.
[6] Lam TT,Wang J,Shen YY,et al. The Genesis and source of the H7N9 influenza viruses causing human infections in China[J].Nature,2013,502(7470):241-244.
[7] Li XY,Shi JZ,Guo J,et al. Genetics,receptor binding property,and transmissibility in mammals of naturally isolated H9N2 avian influenza viruses[J]. PLo S Pathogens,2014,10(11):e1004508.
[8]白洁,刘玉良,关婕葳,等.影响禽流感病毒宿主嗜性和致病性的关键氨基酸位点的研究进展[J].黑龙江畜牧兽医,2018(7):42-45,52.
[9]陈琳. H9N2亚型禽流感病毒的分离鉴定及其遗传进化分析[D].福州:福建农林大学,2015.
[10]何军,刘丽萍,候赛,等.安徽省2株人感染H9N2流感病毒基因特征[J].中华流行病学杂志,2016,37(5):708-713.
[11]徐丹雯. H9N2亚型禽流感病毒的序列分析及其生物学特性研究[D].扬州:扬州大学,2016.
[12]叶贺佳,梁昭平,彭特,等. 2012-2015年H9N2亚型AIV分离株HA基因的克隆及序列分析[J].中国畜牧兽医,2016,43(5):1148-1155.
[13] Xu KM,Smith GJD,Bahl J,et al. The Genesis and evolution of H9N2 influenza viruses in poultry from southern China,2000 to2005[J]. Journal of Virology,2007,81(19):10389-10401.
[14]卢受昇,高慧敏,邓国东,等.广东地区近2年禽流感H_9亚型HA基因的序列分析[J].广东畜牧兽医科技,2014,39(6):29-32,41.