2010年广东省深圳市首例手足口病死亡病例的病原学研究

目的 对广东省深圳市2010年首例临床诊断为手足口病死亡患儿的粪便和咽拭子标本进行病原体型别鉴定,从而追踪其病原的可能来源。 方法 先用实时荧光定量反转录-聚合酶链反应(rRT-PCR)方法检测病毒核酸,再用细胞培养方法从粪便和咽拭子标本中分离病毒,并将分离到的毒株提取核酸后,用普通RT-PCR方法分别扩增肠道病毒71型(EV71)和柯萨奇病毒A组16型(Cox A16)的VP1区全长基因并进行序列测定,序列测定结果与不同国家和地区的肠道病毒株A、B、C基因型代表株进行同源性比较和进化树分析。 结果 该病例的2份标本均为EV71核酸阳性,Cox A16核酸阴性;毒株基因经过核苷酸和氨基酸同源性分析,确定该病例病原体为EV71 C4亚型。 结论 2010年深圳市首例手足口病死亡病例的病原体为EV71的C4亚型。

Etiology of the first death case of hand,foot and mouth disease in Shenzhen,Guangdong,2010

Objective To identify the pathogen of the first clinical death case of hand,foot and mouth disease(HFMD) in Shenzhen in 2010 and find the infection source.Methods Fluorescent RT-PCR was conducted to detect viral nucleic acid of the virus,and virus strains were isolated from stool and throat swabs of the case,then conventional RT-PCR was conducted to amplify full length genes of EV71 and Cox A16 in VPI region of nucleic acid extracted from isolated strains.The gene sequencing,comparison of homology with the representative strains of enteroviruses(genotype A,B,C) isolated in different countries and phylogenetic tree analysis were carried out.Results The two swabs of cases were all positive for EV71 but negative for Cox A16.The gene sequence analysis indicated that the virus isolated belonged to C4 subtype of EV71.Conclusion The pathogen of the first death case of HFMD in Shenzhen was EV71 C4.