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目的对天津市2012年的手足口病病例进行病原学检测,并对引起HFMD流行的肠道EV71型(EV71)和柯萨奇A组16型(CVA16)病毒进行分子生物学特征分析。方法利用Real-time PCR检测HFMD疑似病例标本,用人横纹肌肉瘤(RD)细胞对Real-time PCR检测阳性的部分病例标本进行病毒分离。随机挑取9株EV71病毒分离株和8株CVA16病毒分离株,采用PCR扩增VP1全基因,并进行核苷酸序列和遗传进化分析。结果共检测HFMD疑似病例1 829例,肠道病毒总阳性率为81.30%(1487/1829),其中EV71占39.88%,CVA16占38.33%,其他EV占21.79%。9株EV71分离株与C4亚型的C4a分支代表株的核苷酸序列同源性为97.3%~98.5%,均归属于C4a分支;8株CVA16分离株与B1亚型代表株的核苷酸序列同源性为92.9%~97.1%,其中7株属于B1b分支,1株属于B1a分支。结论导致天津市2012年HFMD流行的EV71为C4亚型中的C4a病毒株,CVA16为B1亚型的B1b病毒株。
Objective To analyze the etiology of hand, foot and mouth disease (HFMD) in 2012 and analyze the molecular biological characteristics of EV71 (EV71) and Coxsackie A (type 16) (CVA16) viruses. Methods Real-time PCR was used to detect the suspected HFMD cases and the rhabdomyosarcoma (RD) cells were used to isolate the virus from the positive cases detected by Real-time PCR. Nine EV71 isolates and eight CVA16 isolates were randomly selected. The whole VP1 gene was amplified by PCR and analyzed by nucleotide sequence and genetic evolution. Results A total of 1 829 suspected cases of HFMD were detected. The total positive rate of enterovirus was 81.30% (1487/1829), of which 39.78% were EV71, 38.33% were CVA16 and 21.79% were other EV. The nucleotide sequence homologies of 9 strains of EV71 isolates and C4 subtype C4a strains were 97.3% ~ 98.5%, belonging to the C4a branch. Among the 8 isolates of CVA16 and B1 subtype, Sequence homology was 92.9% ~ 97.1%, of which 7 belonged to B1b and 1 belonged to B1a. Conclusions The EV71 prevalence in HFMD in Tianjin in 2012 was C4a in C4 subtype and B1 in B1 subtype was CVA16.