【作者】 林君芬； 严菊英； 陈劲华； 孙继民； 张严峻； 傅涛； 茅海燕； 凌峰；

【Author】 LIN Jun-fen,YAN Ju-ying,CHEN Jin-hua,SUN Ji-min,ZHANG Yan-jun FU Tao~2,MAO Hai-yan~1,LING Feng~1.1.Zhejiang Center for Disease Control and Prevention,Hangzhou 310051,China;2.Yiwu Center for Disease Control and Prevention,Yiwu 322000,China.

【机构】 浙江省疾病预防控制中心； 义乌市疾病预防控制中心；

【摘要】 目的通过对病毒分离株的分子特征研究,追踪本次暴发疫情传染来源,为及时有效控制疫情提供科学依据。方法采集疑似病例血清,用荧光RT-PCR方法检测核酸,对核酸检测阳性者进行病毒分离,对分离株提取核酸,再用RT-PCR方法扩增E基因和3’非编码区(3’UTR),测序后经BLAST比对分析其同源性,并用MEGA4.1构建遗传树进行遗传进化分析。结果 40份疑似病人血清中26份荧光RT-PCR检测阳性,分离出23株病毒,分离株的E基因序列完全一致,与其同源性最高的毒株为2004年沙特阿拉伯分离株(D3/Sandi Arabia/2004),为99.3%,与原型株(D3/PH/H87/1956)的同源性为93.4%;分离株的3’-UTR区序列也完全一致,与其同源性最高的毒株为2003年印度分离株(D3/India/GWL-25/2003),为99.1%,与原型株(D3/PH/H87/1956)的同源性为95.8%。E基因和3’-UTR区分别构建的进化树显示浙江登革分离株与沙特阿拉伯和印度株(D3/Sandi Arabia/2004和D3/India/GWL-25/2003)亲缘关系最近,均在登革3型G基因亚型Clade3进化支上,属于印度次大陆型。结论这是一起登革3型GⅢ基因亚型病毒引起的暴发疫情,分离株的分子特征显示本次疫情很可能是由阿拉伯或印度地区输入病例引起。更多还原

【Abstract】 Objective To analyze the molecular characteristic of Dengue virus strains from the outbreak, which will provide scientific information for outbreak control.Methods Serum samples from suspected patients were detected using real-time RT-PCR after being collected.Virus strains were isolated from positive samples.The viral RNAs of the isolated virus strains were extracted and followed by RTPCR to amplify E gene and 3’UTR,which were then sequenced,and the phylogenetic and homology trees were also constructed using MEGA4.1.Results Dengue virus RNAs were detected in 26(65%) serum samples and 23 virus strains were isolated.The sequences of isolated strains E gene were identical and had 99.3%similarity to the sequence of the strain(D3/Sandi Arabia/2004) from Sandi Arabia.The sequences of isolated strains 3’-UTR were also identical and had 99.1%similarity to the sequence of the strain(D3/India/GWL-25/2003) from India.The sequences of E gene and 3’-UTR had 93.4%and 95.8%similarity to the prototype strain(D3/H87/1956),respectively.Phylogenetic trees indicated that D3/ZJ/2009 had maximum similarity with the D3/Sandi Arabia/2004 and D3/India/GWL-25/2003 strains.They all lied in the D3/GⅢ/clade3 branch,which belonged to Indian Subcontinent.Conclusions Molecular characteristic of virus strains confirmed that this outbreak was caused by type 3 dengue virusⅢgenotype,which probably imported from Sandi Arabia or India.更多还原