CAJ | 학술논문

目的对2007年浙江省采集的蚊虫标本进行病毒分离与鉴定.方法 2007年在浙江省采集蚊虫标本,用金黄地鼠肾细胞(BHK-21)分离病毒,对新分离的病毒进行血清学和分子生物学鉴定.结果采集到4735只蚊虫,分离到2株致BHK-21细胞病变的病毒,经免疫荧光和RT-PCR试验鉴定为乙型脑炎病毒(JEV),利用病毒PrM区段进行基因分型分析,新分离的2株病毒属于基因Ⅰ型JEV.对这2株病毒的E基因区段进行分析,核苷酸和氨基酸的同源性分别为99.0%和98.8%.与疫苗株SA14-14-2相比,核苷酸同源性在87.7%,氨基酸同源性在96.4%以上,共有14个共同的氨基酸位点差异.结论在浙江省采集的蚊虫标本中分离到2株病毒,经鉴定为基因Ⅰ型JEV,是浙江省近年来首次分离.
목적 대2007년절강성채집적문충표본진행병독분리여감정.방법 2007년재절강성채집문충표본,용금황지서신세포(BHK-21)분리병독,대신분리적병독진행혈청학화분자생물학감정.결과 채집도4735지문충,분리도2주치BHK-21세포병변적병독,경면역형광화RT-PCR시험감정위을형뇌염병독(JEV),이용병독PrM구단진행기인분형분석,신분리적2주병독속우기인Ⅰ형JEV.대저2주병독적E기인구단진행분석,핵감산화안기산적동원성분별위99.0%화98.8%.여역묘주SA14-14-2상비,핵감산동원성재87.7%,안기산동원성재96.4%이상,공유14개공동적안기산위점차이.결론 재절강성채집적문충표본중분리도2주병독,경감정위기인Ⅰ형JEV,시절강성근년래수차분리.
Objective To study the situation of arboviruses carried by mosquitoes in Zhejiang province.Methods Mosquitoes were collected from Zhejiang province in 2007.Virus strains were isolated by the inoculation of homogenates of the mosquitoes onto BHK-21 cell line.The isolated strains were identified by serological(IFA)and molecular methods(RT-PCR).Results Two strains were isolated from mosquitoes causing cytopathogenic effect(CPE)in BHK-21 cells.Results from serological tests showed that both of the two strains were positive for the antibody to Japanese encephalitis virus(JEV).PrM and E gene were then cloned and sequenced.Results from the phylogenetic analysis showed that the isolates belonged to genotype Ⅰ JEV while through sequence analysis it showed that the homology of nucleotide sequences and amino acid sequences between the two strains were 99.0% and 98.8% respectively.Compared with the JEV vaccine strain SA14-14-2 and two strains,the homology of nucleotide sequences Was up to 87.7% and homology of amino acid sequences was up to 96.4%.When comparing with the vaccine strain SA14-14-2,there were 14 common amino acid variations in all the two strains.Conclusion Two strains of JEV were isolated from mosquitoes collected in Zhejiang province whiCh was the first isolation of genotype Ⅰ JEV in the province in recent years.