上海市浦东新区引起腹泻的腺病毒分子流行病学特征

Molecular epidemiology characterization analysis of diarrhea caused by adenovirus in Pudong New District of Shanghai

目的了解2015—2016年上海市浦东新区病毒性腹泻感染中腺病毒感染的分子流行病学特征。方法采集浦东新区14家二、三级医疗机构肠道门诊中的腹泻患者粪便标本和信息,通过实时荧光聚合酶链式反应(real-time PCR)方法对样本中的腺病毒等5种腹泻类病毒进行初筛;对腺病毒初筛阳性标本,通过人喉癌上皮细胞(Hep-2细胞系)分离腺病毒毒株;通过PCR,扩增腺病毒六邻体蛋白(Hexon蛋白)部分片段并测序,测序结果使用BioEdit软件进行整理,MEGA软件构建腺病毒系统进化树,BEAST软件对腺病毒核苷酸替换率进行分析。结果 2015—2016年,浦东新区疾病预防控制中心共采集腹泻病例样本4 089份,在1 595份腹泻病毒初筛阳性样本中,有腺病毒阳性146份,腺病毒的检出率为3.57%(146/4 089),腺病毒在腹泻病毒中的构成比为9.15%(146/1 595)。腺病毒感染的男女比为1.56∶1,0～9岁是所有年龄组中最易感染腺病毒腹泻的年龄组。腺病毒感染没有明显的季节分布。分离到的58株毒株共有11个腺病毒血清型,以腺病毒41型(22株)和3型(18株)血清型为主。腺病毒的核苷酸变异率为3.53×10^(-6)核苷酸替换/位点/年。结论腺病毒41和3型是本地区腺病毒腹泻的主要病毒基因型,存在腺病毒核苷酸替换,需要加强腺病毒感染腹泻的研究与防控,避免因腺病毒感染导致的聚集性疫情的发生。

[Objective]To understand the molecular epidemiological characterization of diarrhea caused by adenovirus(ADV) in Pudong new area,Shanghai from 2015-2016.[Methods]Fetal samples and information of diarrhea patients were collected from intestinal outpatient clinics of 14 secondary or tertiary medical institutions in Pudong new area. Real-time polymerase chain reaction(real-time PCR)was used to conduct preliminary screening of 5 diarrhea viroid. After screening,selected the ADV positive specimens,cultured by human laryngeal carcinoma epithelial Cells(Hep-2),than conducted PCR to enlarge and sequence the ADV hexaneighbour protein gene(Hexon gene) fragment. The sequencing results were sorted by BioEdit software,and the phylogenetic tree of adenovirus system was constructed by MEGA software. Molecular evolution of adenovirus was analyzed by Bayesian Evolutionary.[Results]Totally 4 089 samples were collected from Pudong new area center for disease control and prevention in2015-2016,of which,1 595 samples were positive for diarrhea viruses and 146 samples were positive for ADV,with a 3.57%(146/4 089)of detection rate. During the surveillance time,9.15%(146/1 595)diarrhea were due to ADV,and the sex ratio was 1.56∶1. Children who were 0-9 years were easily to be infected by ADV,with no obvious seasonal distribution. There were 11 serotypes isolated from58 cultured ADV strains,serotype 41 and 3 were the main serotypes. The variation rate of ADV nucleotide was 3.53×10-6 substitution/site/year.[Conclusion]The main genotypes of ADV diarrhea in this region are serotype 41 and 3,with the presence of nucleotide substitution. It is necessary to strengthen the research,as well as the prevention and control of ADV-infected diarrhea,to prevent the outbreak and aggregate epidemics of ADV infection.