我国广东、福建地区2000～2001年手足口病肠道病毒71型分离株的种系进化分析  

Phylogenetic analysis of enterovirus 71 isolated from patients with hand, foot and mouth disease in Guangdong and Fujian provinces, 2000-2001

作　　者：林思恩[1] 章青[1] 谢华萍[1] 谢健萍[2] 何家鑫[3] 董巧丽[1] 方肇寅[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所腹泻室,北京100052 [2]广州儿童医院 [3]福建省卫生防疫站

出　　处：《中华实验和临床病毒学杂志》2004年第3期227-229,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：美国NIH课题 (R0 3TWO1192 );国家自然科学基金资助 ( 3 0 2 70 0 69)

摘　　要：目的　采用逆转录聚合酶链反应 (RT PCR)检测我国广东、福建地区 2 0 0 0～ 2 0 0 1年手足口病 (HFMD)散发病例中的肠道病毒 71型 (EV71) ,进而通过扩增VP1节段的核苷酸序列 ,进行毒株的种系进化分析。方法　以肠道病毒特异引物对EV 1、EV 2进行RT PCR ,经琼脂糖凝胶电泳鉴定为阳性的标本 ,进一步用EV71特异引物对 15 9S、16 2A进行RT PCR ,扩增的VP1节段经纯化后与测序载体pGEM T连接 ,转化大肠埃希菌DH5α ,筛选后测序。所得序列与我国深圳、上海、武汉地区的EV71流行株 ,中国台湾 1998年 4例HFMD暴发分离的EV71毒株 ,及来自美国、日本、匈牙利等国家地区的EV71毒株的核苷酸序列 ,用ClustalX 1 8和PHYLIP 3 5进行比对分析 ,构建种系进化树。结果　 2 5份标本检测EV71的阳性率为 2 0 % ,所得序列经种系进化分析 ,与肠道病毒 71型的其他毒株同源 ,与深圳 1998年HFMD散发分离的EV71毒株同源性为 94 % ,与上海 2 0 0 0年HFMD暴发分离株同源性为94 %～ 96 % ,与武汉 1987年HFMD病例中分离的毒株同源性为 91% ,而与国外EV71毒株同源性仅为82 %～ 84 %。结论　EV71是我国南方地区HFMD的主要病原之一 ;我国大陆地区的EV71毒株在种系进化上有较高的同源性 ;与我国台湾地区大部分分离株亦有 90 %～ 91%的核苷酸?Objective To identify enterovirus 71 (EV71) strains isolated from patients with hand, foot and mouth disease (HFMD) in Guangdong and Fujian provinces from 2000 to 2001 by using phylogenetic analysis Methods All 25 samples were first tested for enteric viruses by RT-PCR using enterovirus specific primers EV-1 and EV-2, and then were identified for EV71 by RT-PCR using EV71 specific primers 159S and 162A The amplicons of 485bp segment (part of the VP1 gene) were cloned into pGEM-T and sequenced A phylogenetic tree was constructed by comparison of the sequences with other 12 EV71 strains isolated from China, Japan, Hungary, and the United States including the prototype BrCr Results The positive rate of EV71 was about 20% The sequence analysis showed that the new isolate (GZH2000) shared 94%-96% nucleotide identity with three strains isolated in 1998 and 2000, and 91% with a strain isolated in 1987 from Chinese mainland, but shared only 82%- 84% homology with EV71 isolates studied abroad Conclusion EV71 is one of the important pathogens of HFMD in south ChinaThe strains isolated from mainland were closely related with most isolates from Taiwan, but different from most EV71 strains reported abroad The symptoms of EV71 infection in mainland were not as intensive as those described in Taiwan's outbreak

关 键 词：HFMD 肠道病毒71型 种系 手足口病 毒株 暴发 RT-PCR 进化分析 同源性 分离株 

分 类 号：R373[医药卫生—病原生物学] R725[医药卫生—基础医学]