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目的分析云南省腾冲县新分离流行性乙型脑炎病毒(JEV)和盖塔病毒(GETV)的分子生物学特征。方法2007年7月在中缅边境地区腾冲县采集蚊虫标本,用BHK-21和C6/36细胞分离病毒,阳性分离物用RT-PCR方法进行鉴定,用相关生物学软件进行分子生物学特征分析。结果从腾冲县采集的4属25种26 702只蚊虫中分离到20株JEV,1株GETV。系统进化分析表明,新分离20株JEV均属于基因Ⅰ型,与邻近省份和东南亚流行株亲缘关系较近;PreM和E基因核苷酸同源性和氨基酸位点分析表明,这20株JEV之间及其与国内外流行株间均存在一定差异,但它们的抗原和毒力位点未发生明显改变。本次分离的GETV的Capsid基因和E2基因与其他国内外分离株的同源性较高。结论首次证实云南省西部地区存在基因Ⅰ型JEV和GETV流行,这些流行株具有较为稳定的遗传特征和地域特征。
Objective To analyze the molecular characteristics of newly isolated Japanese encephalitis virus (JEV) and captive virus (GETV) in Tengchong County, Yunnan Province. Methods In July 2007, mosquitoes were collected from Tengchong County in the Sino-Burmese border area. Viruses were isolated from BHK-21 and C6 / 36 cells. The positive isolates were identified by RT-PCR and analyzed by molecular biology . Results 20 JEV and 1 GETV were isolated from 25 mosquitoes of 26 702 mosquitoes in 25 genera collected from Tengchong County. Phylogenetic analysis showed that all of the 20 new JEV isolates belonged to genotype Ⅰ, which were closely related to the epidemic strains in neighboring provinces and Southeast Asia. The nucleotide homology and amino acid sequence analysis of PreM and E showed that these 20 JEV There are some differences between the strains and the epidemic strains at home and abroad, but their antigen and virulence sites have not changed significantly. The Capsid gene and E2 gene of GETV isolated this time have high homology with other isolates from other countries. Conclusion It is the first time to confirm that there is genotype Ⅰ JEV and GETV epidemic in western Yunnan. These epidemic strains have relatively stable genetic and geographical features.