论文部分内容阅读
目的了解河南省部分地区蚊传虫媒病毒的种类分布及基因型别。方法于2012年5-8月在河南省洛阳市新安县和信阳市息县的居民住房、猪圈、牛棚及树林中采集蚊虫。使用细胞培养法分离病毒,通过反转录-聚合酶链反应(RT-PCR)方法进行种属特异扩增鉴定,并用Clustal X2.1、Meg Align、Genedoc 3.2和Mega v5.1生物信息学软件完成病毒核酸序列的分子生物学分析并进行基因分型。结果共采集蚊虫4属5种7149只,其中新安县以骚扰阿蚊居多(2055只,51.36%),息县以淡色库蚊(2964只,94.16%)为主。共分离获得5株病毒,直接接种BHK-21细胞无病变、C6/36细胞培养仅导致轻微病变,其C6/36细胞传代培养物转接BHK-21细胞后3 d可致细胞明显病变;序列分析表明这5株病毒均属基因Ⅰ型流行性乙型脑炎病毒(JEV),其中3株分离自淡色库蚊,2株来自三带喙库蚊。结论河南省部分地区依然存在JEV及其传播媒介,且目前当地自然界中循环的JEV仍以基因Ⅰ型为主。
Objective To understand the species distribution and genotype of mosquito-borne Arbovirus in some areas of Henan Province. Methods From May to August 2012, mosquitoes were collected from residential buildings, pigsty, bullpen and woods in Xin’an County, Luoyang City, Henan Province and Xixian County. Viruses were isolated using cell culture and species-specific amplification by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and identified using Clustal X2.1, Meg Align, Genedoc 3.2 and Mega v5.1 bioinformatics software Molecular biology analysis of the viral nucleic acid sequence is completed and genotyped. Results A total of 7149 mosquitoes belonging to 4 genera and 5 species were collected. Among them, Xin’an County mainly contained mosquitoes (2055, 51.36%), and Culex pipiens pallens (2964, 94.16%) in Xixian County. Five strains were isolated and directly inoculated with BHK-21 cells without any pathological changes. C6 / 36 cell culture only resulted in slight pathological changes. The passage of BHK-21 cells from C6 / 36 cell subcultured cells resulted in obvious pathological changes. Analysis showed that all of the five viruses belonged to genotype I Japanese encephalitis virus (JEV), of which three were isolated from Culex pipiens pallens and two from Culex tritaeniorhynchus. Conclusion JEV and its transmission media still exist in some areas of Henan Province. At present, the circulating type JEV in the local nature is still dominated by genotype Ⅰ.