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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。
Objective To identify the pathogen types of stool and throat swab specimens from the first case of death from HFMD in Shenzhen City, Guangdong Province in 2010, so as to trace the possible origin of the pathogen. Methods The viral nucleic acid was detected by real-time quantitative reverse transcription-polymerase chain reaction (rRT-PCR), then the virus was isolated from the stool and throat swab samples by cell culture and the isolated strains were extracted nucleic acid . The full-length VP1 gene of enterovirus 71 (EV71) and Cox A16 (Cox A16) was amplified by the ordinary RT-PCR method and sequenced. The results of sequencing were consistent with those of different countries and regions The homology comparison and phylogenetic tree analysis of representative strains of A, B, C genotypes of enterovirus strains. Results The two samples of this case were positive for EV71 and negative for Cox A16. The nucleotide and amino acid homology analysis showed that the pathogen was EV71 C4 subtype. Conclusion The first case of death from HFMD in Shenzhen in 2010 was the C4 subtype of EV71.