辽宁省啮齿类动物肾综合征出血热病毒株的分离及鉴定

Isolating and identifying hemorrhagic fever with renal syndrome vivus from rodent animal in Liaoning Province

目的:通过对我省疫区捕获的鼠肺样本做病毒分离,初步掌握辽宁省啮齿类动物自然感染肾综合征出血热(HFRS)病毒亦称为汉坦病毒(Hantavirus,HV)的病毒型别,为今后疫区的防治工作提供可靠的依据。方法:将待检的阳性鼠肺样本分成二组,其中一组直接用vero-E6细胞做病毒分离,而另一组先做动物试验将病毒增殖,然后再做vero-E6细胞传代适应;最后应用型特异性引物进行逆转录套式PCR(RT-PCR,)扩增病毒核酸,并对其M片段的靶序列核苷酸序列进行测定,从而达到对所分离的汉坦病毒进行分型及鉴定。结果:样本直接做细胞分离获得阳性病毒株3株,经动物增殖两代后获得强阳性鼠肺3份,弱阳性鼠肺1份;再转vero-E6细胞传代适应后得到阳性毒株2株,经核酸检测与序列鉴定确定了我们所分离的5株病毒株均为汉坦病毒,其中1株为汉滩型病毒(Hantaan virus,HTNV),4株为汉城型病毒(Seoul virus,SEOV)。

Objective：We initially learned the type of hemorrhagic fever with renal syndrome HFRS virus （Hantavirus）, which rodent were naturally infected in Liaoning province, by separating virus from lungs of mice in HFRS epidemic area. And it will provide reliable bases for the work of prevention and control. Methods： We divided the subjects of mice lung into two groups ： in the first group the virus was separated directly by vero - E6 cells and in the second one it was firstly multiplied in vivo and separated in the same way; Then we used RT - PCR to amplify RNA of two groups virus above by type specific primer and sequenced the M piece in it. Results：We separated 3 strains of virus in the first group, while from the second group animals we received 3 portions of strong positive subjects and 1 portion of light one, from which we got 2 strains of virus. We identified them as Hantavirus, one was Hataan virus （HTNV） , the other four were Seoul virus （SEOV）.