河北省2008年手足口病病毒的分离和鉴定

Isolation and identification of viruses from patient with hand,foot and mouth disease in Hebei Provice in 2008

目的：了解引起河北省2008年5-8月手足口病（HFMD）流行的病原体。方法：将HFMD患者的脑脊液、疱疹液、粪便、咽拭子四种标本接种RD细胞进行病毒分离,用肠道病毒基因的特异性通用引物序列、肠道病毒71型（Entero-virus71,EV71）和柯萨奇A组16型（Coxsackivirus16,CA16）基因的特异性引物序列进行RT-PCR鉴定;采用微量板细胞病变法（CPE）测定半数细胞感染浓度。结果：2008年5-8月间从203份手足口病例标本（脑脊液9份,疱疹液27份,粪便91份,咽拭子76份）中分离出115株病毒,病毒分离阳性率为56.65%,分离到的病毒经RT-PCR鉴定,其中72株为EV71,42株为EV71＋CA16,1株为非EV71、非CA16其他肠道病毒,各标本之间病毒分离阳性率的差异无统计学意义（P〉0.05）;粪便标本的病毒滴度高于疱疹液和咽拭子标本。结论：河北省2008年5-8月HFMD流行的原因主要由EV71感染所引起。

Objective ：To detect the etiological agent of hand, foot and mouth disease（HFMD） epidemics in Hebei provice from May to August of 2008. Methods： The specimens（ including cerebral spinal fluid, vesicle fluid, stool and throat swabs ） were inculated into RD cells for isolation of enteroviruses（EV）. The isolates were identificated for EV, Enterovirus71 （Enterovirus71, EV71 ） and Coxsackivirus16（CA16） using revers transcription polymerase chain reaction（ RT - PCR） ; 50% cell culture infective dose （CCIDso） was based cytopathic effect test（CPE）. Results：Total 115 viruses were isolated from 203 specimens（56. 65% ）, of which 72 were EV71,42 were EV71 ＋ CA16 and 1 was were Non -EV71 ,CA16 EV. The viruses isolation rate was not satistcally significant in different specimens. The virus titration of stools was higher than that of vesicle fluid and throat swabs. Conclusion：EV71 is the main cause of HFMD during the epidemic.