摘要
目的对深圳市2005年从登革热患者急性期血液中分离到的1株登革病毒进行型别鉴定,从分子水平分析分离株的生物学特征,追踪其可能的地域来源。方法用C6/36细胞培养增殖病毒株SZ0524,收集病毒液。用逆转录-半套式PCR(RT-semi-nested-PCR)方法和荧光PCR方法对其进行型别鉴定。扩增病毒E基因后进行序列测定,并与不同国家和地区的登革病毒株进行同源性比较和进化树分析。结果深圳市登革病毒分离株用4型特异性引物扩增出392bp的特异性条带,荧光PCR进一步证实了分离的病毒株为登革4型病毒。SZ0524与登革病毒4型国际标准株H241株在E基因上核苷酸同源性为99.7%,而与登革病毒1、2、3型国际标准株HAWAII、NGC、H87同源性分别为57.0%、59.2%和56.2%。基因进化树显示SZ0524株与D4-73NIID株和D4-61NIID株亲缘关系最近,其次为H241,在进化树的同一分支上,属基因Ⅰ亚型。结论从分子水平证明从深圳市登革热患者血清中分离到的毒株确为DEN-4型病毒。结合流行病学调查资料,证实此病例为输入性感染病例,该毒株最有可能来源于东南亚一带。
To identify the genotype and analyze the molecular characteristics of dengue virus strain SZ0524 isolated from serum samples of patients with early stage of dengue fever in Shenzhen in 2005 so as to explore its possible origin.The C6/36 cell line was cultivated with virus strain SZ0524 and its suspension was harvested.The type of isolated virus strain was determined by RT-semi-nested PCR and fluorescent PCR.E gene of isolated virus strain was amplified by RT-PCR and sequenced.Homology and phylogenetic tree of E gene of this dengue virus with the strains isolated from other areas were constructed.This SZ0524 strain was further identified by fluorescent PCR,and confirmed to be the type 4 virus after obtaining the 392bp band with type 4 specific primers.The homology of nucleotide sequence of E gene of SZ0524 strain with the standard type 4 dengue virus H241 strain were 99.7%,but the homology with the standard dengue virus 1,2,3 in the same fragment were 57.0%,59.2% and 56.2% respectively.Analysis of the phylogenetic tree indicated that SZ0524 was more close to D4-73NIID and D4-61NIID strain,next to H241 strain,and they lied in the same branch of phylogenetic tree.The isolated dengue virus type 4 belonged to genotype Ⅰand the SZ0524 strain was proved to be dengue virus type 4 in the molecular level.Combined with epidemiology information,it is suggested that this case can be classified as an imported case and the SZ0524 strain may be transferred from the southeast asian region.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2010年第1期17-20,共4页
Chinese Journal of Zoonoses
