摘要目的 研究2010年南京市手足口病的病原构成及肠道病毒71型(EV71) VP1编码区基因特征.方法 采集南京儿童医院手足口病住院患儿的248份咽拭子标本,进行病毒分离,然后利用real-time PCR法对阳性分离物进行肠道病毒属、EV71和柯萨奇病毒A16型(Cox A16)鉴定.选取临床诊断为普通型病例、重型病例和危重型病例的EV71共20株进行VP1编码基因区基因扩增及核苷酸序列测定和分析,然后与EV71各基因型和基因亚型的代表株构建亲缘性进化树.结果 248份咽拭子标本共分离出146株病毒分离物.其中110株EV71,阳性率为44.35％,28株Cox A16,阳性率为11.29％,其他肠道病毒8株,占3.23％.对20株EV71进行VP1区核苷酸和氨基酸分析,发现其同源性较高,分别为95.51％ ～ 100％和98.32％ ～ 100％.构建亲缘性进化树显示20株EV71与C4基因亚型的代表株处于同一分支,属于C4基因亚型.结论 南京地区2010年手足口病病原以EV71为主,且流行基因型为C4基因亚型；不同病例分型的EV71在VP1区未发现明显差异.

abstractsObjective To study the etiological agent of hand-foot-and-mouth disease ( HFMD),and to genetically characterize enterovirus 71 ( EV71 ) isolated from clinical specimens of HFMD patients in Nanjing in 2010. Method Throat swab specimens were collected from 248 inpatients with HFMD in Nanjing Children's Hospital and then viral isolation was performed.Real-time PCR was used for detection of EV71,coxsaekievirus A16 (Cox A16) and other enteroviruses from the positive isolates. Twenty EV71 strains from different clinical types of cases were selected for entire VP1 coding gene amplification and sequencing,finally a phylogenetic tree was constructed among the 20 EV71 strains and EV71 representative strains of known genotypes and subgenotypes. Result From the 248 throat swabs specimens,110 EV71 strains,28 Cox A16 strains,and 8 other enterovirus strains were isolated and the positive rate was 44.35％,11.29％,3.23％,respectively.Then nucleotide sequencing was performed on the 20 EV71 strains.There was little difference in the nucleotide and the amino acid sequences among the 20 EV71 strains,the homology was 95.51％ -100％ and 98.32％ -100％,respectively.The phylogenetic tree showed that all the 20 EV71 strains belonged to C4 subgenotype. Conclusion EV71 was the main pathogen of HFMD in Nanjing in 2010 and all the analyzed isolates belonged to C4 serotype.No significant difference was found in sequence of VP1 region of EV71 isolated from different clinical types of cases.