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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: To isolate and isolate viruses from murine lung samples captured in the epidemic area in our province and to master the virus type of Hantavirus (HV) naturally infected with rodent hemorrhagic fever with renal syndrome (HFRS) virus in Liaoning province. , To provide a reliable basis for the prevention and control of epidemic areas in the future. Methods: The positive rat lung samples to be tested were divided into two groups, one of them directly used vero-E6 cells for virus isolation, while the other group used animal experiments to proliferate the virus and then passed through the vero-E6 cells. Finally, Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify the viral nucleic acid and the nucleotide sequence of its target M fragment was determined. The typing of the isolated Hantavirus Identification. Results: Three strains of positive virus were obtained by direct cell isolation. Three passages of strong positive rat lung and one weakly positive rat lung were obtained after two generations of animal multiplication. Two strains of vero-E6 were passaged and passaged again The five strains of Hantavirus were identified as Hantaan virus (HTNV) and four of them were Seoul virus (SEOV) by nucleic acid detection and sequence identification. .