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目的对上海市2008～2009年手足口病(Hand,Footand Mouth Disease:HFMD)部分病例,进行病原学与分子流行病学研究。方法用人横纹肌肉瘤(Human Rhabdomyosarcoma,RD)细胞和非洲绿猴肾(African Green Monkey Kidney,Vero)细胞,对采集到的HFMD病例临床标本进行病毒分离,用逆转录-聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)、序列测定和同源性分析的方法,对分离到的病毒进行分析鉴定。结果上海市2008～2009年分别从66.5%(105例)和69.0%(20例)的患者中分离到肠道病毒(Enterovirus,EV),分离到的EV71型(EV71)属于C4基因亚型,但与上海市2002年EV71分离株相比已发生了较大的变异。分离到的柯萨奇病毒A组16型(Coxsackievirus Group A Type16,CA16)属于B基因型,分成2个明显的分支,每个分支内部都有近两年的病毒共存,且与上海市2002年CA16分离株相比变异不大。结论 2008～2009年导致上海市HFMD流行的病毒仍以EV71和CA16为主;在采用RD和Vero两种细胞同时分离病毒时,不同类型标本的病毒分离阳性率差异无统计学意义。
Objective To investigate the etiology and molecular epidemiology of some cases of Hand, Foot and Mouth Disease (HFMD) in Shanghai from 2008 to 2009. Methods Human Rhabdomyosarcoma (RD) cells and African Green Monkey Kidney (Vero) cells were used to isolate the virus samples from clinical samples collected from patients with HFMD. Reverse Transcription- Polymerase Chain Reaction, RT-PCR), sequence analysis and homology analysis, the isolated virus was analyzed and identified. Results Enterovirus (EV) was isolated from 66.5% (105 cases) and 69.0% (20 cases) patients in Shanghai from 2008 to 2009. The isolated EV71 (EV71) belonged to C4 subtype, However, a large variation has occurred in comparison with the 2002 EV71 isolate in Shanghai. The isolated Coxsackievirus Group A Type 16 (CA16) belongs to the B genotype and is divided into two distinct branches, each of which has a virus coexistence within the past two years and is in close cooperation with Shanghai 2002 CA16 isolates showed little variation. Conclusions The viruses that caused the epidemic of HFMD in Shanghai from 2008 to 2009 were still mainly EV71 and CA16. There was no significant difference in the positive rates of virus isolation between different types of samples when virus was separated by RD and Vero cells simultaneously.