标题：2016年上海市普陀区手足口病病原谱及EV71和Cox A16病毒分离株VP1基因特征分析

      作者：李晓君，唐海丰，何鑫，胡焰，顾文超    (上海市普陀区疾病预防控制中心，上海 200333)

      卷次： 2018年29卷20期

      【摘要】 目的 了解上海市普陀区手足口病病原学特点，分析肠道病毒71型(EV71)和柯萨奇病毒A组16型(Cox A16) VP1基因的特征，为手足口病的防治提供科学依据。方法 收集2016年上海市普陀区手足口病患者的咽拭子标本，采用实时荧光定量 PCR法(Real-time PCR)进行核酸检测，并用 RD细胞对核酸检测结果为 EV71和Cox A16的标本进行病毒分离，收获的阳性分离株利用逆转录-聚合酶链反应(RT-PCR)法扩增病毒 VP1区核苷酸序列，测序后分析其同源性并构建系统进化树。结果 2016年上海市普陀区共检测234份手足口病标本，阳性标本138份，阳性率为58.97% (138/234)。其中 EV71阳性率为11.54% (27/234)、Cox A16阳性率为18.80% (44/234)、Cox A6阳性率为 27.78% (65/234)和其他肠道病毒阳性率 0.85% (2/234)。病毒分离培养阳性株包括 10株 EV71和 20株Cox A16，10株 EV71分离株均为 C4a型，VP1区核苷酸和氨基酸序列同源性分别为93.6%~99.9%和98.7%~100.0%；20株 Cox A16分离株均为 B1b亚型，VP1区核苷酸和氨基酸序列同源性分别为 93.4%~100.0%和 98.2%~100.0%。结论 2016年上海市普陀区手足口病呈现 Cox A6、EV71和 Cox A16共同流行，而 EV71流行株属于C4a亚型，Cox A16流行株属于 B1b亚型，均与国内大部分地方的流行株基因亚型基本一致。
      【关键词】 手足口病；病原谱；肠道病毒71型；柯萨奇病毒A组16型；VP1基因
      【中图分类号】 R373.2 【文献标识码】 C 【文章编号】 1003—6350（2018）20—2911—04

Pathogen spectrum of hand-foot-and-mouth disease and genetic characteristics of VP1 from EV71 and Cox A16in Putuo District of Shanghai, 2016.
LI Xiao-jun, TANG Hai-feng, HE Xin, HU YAN, GU Wen-chao. Shanghai PutuoCenter for Disease Control and Prevention, Shanghai 200333, CHINA
【Abstract】 Objective To understand the pathogenic features of hand-foot-and-mouth disease (HFMD) in PutuoDistrict of Shanghai and analyze the genetic characteristics of VP1 from enterovirus type 71 (EV71) and Coxsackie virusA16 (Cox A16), so as to provide scientific evidence for the prevention and treatment of HFMD. Methods Nasopharyn-geal swabs were collected from HFMD patients in Putuo District of Shanghai in 2016, and the viral RNA detection wasperformed by real-time fluorescent PCR (RT-PCR). Viral isolation was performed with rhabdomyosarcoma (RD) cell onthe specimens with the positive RNA detection for EV71 and Cox A16. Entire VPl coding region amplification fromthese positive strains was performed by RT-PCR and sequencing analysis was determined. Sequence analysis and typeidentification and the phylogenetic tree construction were performed. Results Totally 234 HFMD swabs were detected inPutuo District of Shanghai in 2016, and 138 were EV positive, with the positive rate of 58.97% (138/234). The positiverates for EV71, Cox A16, Cox A6 and other enteroviruses were 11.54% (27/234), 18.80% (44/234), 27.78% (65/234) and0.85% (2/234), respectively. A total of 10 EV71 and 20 Cox A16 isolations were obtained, and 10 EV71 strains belongedto sub-genotype C4a. The sequences analysis showed that 10 EV71 strains shared 93.6%-99.9% nucleotide similarityand 98.7%-100.0% amino acid similarity with each other. The 20 Cox A16 strains belonged to sub-genotype B1b. Thesequences analysis showed that 20 CoxA16 strains shared 93.4%-100.0% nucleotide similarity and 99.8%-100.0% ami-no acid similarity with each other. Conclusion The prevalent pathogens of HFMD in Putuo District of Shanghai in2016 are Cox A6, EV71 and Cox A16. The EV71 strains belonged to sub-genotype C4a, and the Cox A16 strains be-longed to sub-genotype B1b. They are closely related with some mainland Chinese isolates.
      【Key words】 Hand-foot-and- mouth disease (HFMD); Pathogen; Enterovirus type 71 (EV71); Coxsackie virusA16 (Cox A16); VP1 gene·疾病监控·doi:10.3969/j.issn.1003-6350.2018.20.031基金项目：上海市普陀区科技计划项目(编号：普KW15313)