摘要
目的分析2015—2018年北京市东城区甲型H1N1pdm09亚型流感病毒的分子流行病学特征。方法收集2015年1月1日—2018年12月31日期间北京市东城区流感样病例的咽拭子样本,RT-PCR法检测核酸,阳性样本接种至MDCK细胞进行病毒分离,血凝抑制试验鉴定流感亚型。选取30株甲型H1N1pdm09亚型分离株,RT-PCR法扩增血凝素(hemagglutinin,HA)和神经氨酸酶(neuraminidase,NA)基因序列,并测序。应用DNASTAR 5.0软件将HA、NA序列进行拼接,MEGA 6.0软件建立系统进化树,并进行遗传特征分析。结果共检测样本7533份,甲型H1N1pdm09亚型231份,阳性率为23.24%。30株甲型H1N1pdm09亚型分离株与WHO推荐的北半球疫苗株A-Brisbane-02-2018的HA基因核苷酸和氨基酸序列同源性分别为97.42%~99.53%和97.40%~99.32%;NA基因核苷酸和氨基酸序列同源性分别为98.11%~99.74%和97.72%~99.80%。HA基因共有16个氨基酸位点发生改变,NA基因共有10个位点发生改变;抗原受体结合位点、糖基化位点和耐药位点均未发生改变。结论2015—2018年北京市东城区甲型H1N1pdm09亚型流感病毒HA和NA基因位点发生了一些变异,但抗原性和耐药性未发生改变,与疫苗株匹配性较高。本研究为北京市东城区评价疫苗保护效果及制定流感防治措施提供了实验依据。
Objective To analyze the molecular epidemiological characteristics of influenza A virus subtype H1N1pdm09 in Dongcheng District,Beijing City,China from 2015 to 2018.Methods Throat swabs were collected from patients with suspected influenza in Dongcheng District,Beijing City from January 1,2015 to December 31,2018 and tested for nucleic acid by RT-PCR,of which the positive ones were inoculated into MDCK cells for virus isolation and identification of influenza subtype by hemagglutination inhibition(HI)test.Thirty isolates of subtype H1N1pdm09 were selected,from which hemagglutinin(HA)and neuraminidase(NA)genes were amplified by RT-PCR,sequenced,and spliced by using DNASTAR 5.0 software.A phylogenetic tree was constructed by using MEGA 6.0 software for analysis of genetic characteristics.Results A total of 7533 samples were detected,from which 231 isolates were of subtype H1N1pdm09,indicating a positive rate of 23.24%.The homologies of nucleotides and amino acids of HA gene of 30 isolates of subtype H1N1pdm09 to those of Northern Hemisphere vaccine strain A-Brisbane-02-2018 recommended by the WHO were97.42%~99.53%and 97.40%~99.32%,while those of NA gene were 98.11%~99.74%and 97.72%~99.80%,respectively.Changes were observed in 16 amino acid sites of HA gene and 10 amino acid sites in NA gene.However,no changes were observed in antigen receptor binding sites,glycosylation sites and drug resistance sites.Conclusion Influenza A virus subtype H1N1pdm09 in Dongcheng District,Beijing City from 2015 to 2018 showed a certain variations in HA and NA gene sites,while antigenicity and drug resistance showed no changes,indicating a high compatibility to the vaccine strain.The study provided an experimental basis for evaluating the protective effect of vaccine and formulating the stratety for influenza prevention and control in Dongcheng District,Beijing City.
作者
李艳宇
李姗姗
杜嘉鑫
韩营营
LI Yan-yu;LI Shan-shan;DU Jia-xin;HAN Ying-ying(Microbiological Laboratory,Dongcheng District Center for Disease Control and Prevention,Bejing Ciy,Beijing 100050,China)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2022年第9期1076-1081,1089,共7页
Chinese Journal of Biologicals
基金
北京市东城区科技计划项目(东卫健研[2019]-11)。
作者简介
通信作者:李艳宇,E-mail:spring200424@126.com。
