目的 分离、鉴定中山市2007—2014年1型登革病毒并获得E基因序列,从分子水平分析其系统进化和地域来源情况,了解该病毒各年份的流行性质.方法 收集中山市2007—2014年登革热患者急性期血清,接种C6/36细胞分离登革病毒,应用RT-PCR技术扩增病毒的包膜蛋白E基因全长,并对序列进行系统进化和同源性分析.结果 共分离到登革1型病毒流行株25株,参考国际标准株E基因序列将其分为GⅠ型和GⅤ型2个基因型.其中2007年3株,2012年3株,2013年3株,2014年7株聚集在GⅠ型中,2014年9株聚集在GⅤ型中.中山2014年登革病毒GⅠ型流行株与中山市2007年、2012年、2013年流行株E基因核苷酸(氨基酸)序列相似度分别为98.25%(99.40%)、98.18%(99.80%)、99.73%(100.00%),与珠三角2014年广州、珠海输入中山株的氨基酸序列相似度为100.00%;中山市2013年登革病毒GⅠ型流行株与广州、佛山的2013年流行株E基因核苷酸(氨基酸)序列相似度分别为99.67%(100.00%)、99.74%(100.00%).结论 2007—2014年中山市流行的登革1型病毒有2个基因型(GⅠ型和GⅤ型).2014年GⅤ型可能由国外输入,GⅠ型中2007年和2012年为国外输入株,2013年及2014年流行株E基因序列相似度高,2014年的登革流行可能与2013年流行株本地化有关.%Objective To obtain the E gene sequences of dengue type 1 viruses isolated from 2007 to 2014 in Zhongshan city,and to analyze their phyletic evolution,geographic origin and epi-demical features.Methods Dengue viruses were isolated in C6/36 cells from the acute phase ser-um of patients with dengue fever from 2007 to 2014.The envelope protein E gene was amplified by RT-PCR,and the homology and phylogeny were analyzed by sequencing.Results A total of 25 dengue type 1 virus strains were isolated,which were identified as two genotypes:GⅠand GⅤ. Among the 25 strains,16 were clustered in genotype GⅠ(3 isolated in 2007,3 in 2012,3 in 2013, and 7 in 2014),and 9 in genotype GⅤ(isolated in 2014).The nucleotide(amino acid)sequence similarities of dengue virus strains isolated in 2014 to those isolated in 2007,2012 and 2013 were 98.25%(99.40%),98.18%(99.80%)and 99.73%(100.00%)in Zhongshan,respectively.Fur-thermore,the amino acid sequence similarities of dengue virus strains isolated in Zhongshan to those imported from Guangzhou and Zhuhai were both 100.00% in 2014.The nucleotide(amino acid)sequence similarities of dengue virus strains isolated in Zhongshan to those isolated from Guangzhou and Foshan were 99.67%(100.00%)and 99.74%(100.00%)in 2013,respectively.Conclusion Dengue type 1 viruses isolated from Zhongshan between 2007 and 2014 were identifiedas two genotypes.Among them,genotype GⅤstrains might be imported from abroad,and genotype GⅠstrains isolated in 2007 and 2012 were foreign strains.The E gene sequences of denguetype 1 viruses showed a high similarity between strains isolated in 2013 and strains isolated in2014,suggesting that dengue epidemic in 2014 may be correlated with strain localization in 2013.
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