广东珠海市2007年登革病毒分子流行病学分析

目的了解2007年珠海市登革热暴发期间登革Ⅰ型（ZH1067/07）及Ⅱ型（ZH1340/07）病毒序列特征和可能的传播来源。方法以GenBank公布的病毒AB178 040和M29 095为参考,分别设计PCR引物,扩增病毒ZH1067/07和ZH1340/07全基因组的不同片段并测序,通过末端重叠序列进行拼接组成病毒全基因组序列。序列GenBank登录号分别为Eu359 008和Eu359 009。参考已公布的登革Ⅰ/Ⅱ型病毒全基因组序列进行进化分析。结果拼接后的登革Ⅰ型病毒基因组全长10 735个核苷酸,经序列进化分析,属登革Ⅰ型中的第1群（基因Ⅰ型）,与2004年分离自日本的登革Ⅰ型毒株AB178 040及分离自福建的登革Ⅰ型毒株Fj231/04核苷酸同源性最高,达99%（10 638/10735）,经流行病学证据支持福建输入病例导致珠海本地暴发;拼接后的登革Ⅱ型病毒基因组全长10 723个核苷酸,经序列的进化分析,属登革Ⅱ型病毒中的第4群（基因Ⅳ型）,与2005年分离自文莱的登革Ⅱ型EU179 857和EU179858两毒株距离最近,核酸同源性99%,（10 609/10 705）。流行病学调查证明2例均为澳门输入病例。结论2007年珠海登革Ⅰ型病毒ZH1 067/07可能来源于福建登革Ⅰ型毒株Fj231/04,与来源于密克罗尼西亚的AB178 040有很近的亲缘关系;珠海登革Ⅱ型病毒ZH1340/07可能由澳门输入,与文莱EU179 857有很近的亲缘关系,未导致珠海本地暴发。

Phylogenetic analysis on dengue viruses in Zhuhai outbreak during 2007

Objective To characterize the Dengue virus type Ⅰ/Ⅱ strain ZH1067/07 and ZH1340/07 in Zhuhai city and explore the possible viral origin.Methods The primers used for amplification and sequencing of Dengue virus type Ⅰ and Dengue virus type Ⅱ genomic sequences were designed separately based on strains AB178040 and M29095.The amplification fragments were sequenced on both strand by using these primers.The sequencing results were assembled into a viral complete genomic sequence by spliced the terminals overlapped each other.The viral genomic sequences were submitted to the GenBank（Accession No： Eu359008 and Eu359009）.The genetic distances were calculated and the phylogenetic trees were constructed by compared the genomic sequences with present sequences available on GenBank.Results The complete genome of Dengue virus type Ⅰ strain of ZH1067/07 was composed of 10735bp and comprised a large open reading frame（ORF）.The phylogenetic analysis demonstrated that it was closest to AB178040 and Fj231/04（99% identity,10638/ 10735）.The epidemic evidence supported that Dengue virus type Ⅰstrain caused outbreak in zhuhai 2007 was originated from Fj231/04.The genom sequence of Dengue virus type Ⅱ strain of ZH1340/07 was composed of 10723bp.It consisted of 96bp and 454bp at The 3＇ terminal and 5＇ terminal non-coding region separately and an open reading frame（ORF）locating in 97~10269 nucleotides.The phylogenetic analysis demonstrated that it was closest to EU179857 and EU179858 strains from Brunei.The epidemic investigation showed that the 2 input cases from Macao were no relationship with outbreak in Zhuhai of 2007.Conclusion Dengue virus type Ⅰ,which genetically closest to Japan strains（AB178040 from Micronesia）,was input from Fujian province of China and caused outbreak in Zhuhai of 2007.Dengue virus type Ⅱ nearest to Brunei strain EU179857,meanwhile inputed cases from Macao,had no relationship with Zhuhai outbreak of 2007.