两株云南省新分离版纳病毒的鉴定及序列分析  

Isolation,identification and sequence analysis of two newly isolated Banna viruses from Yunnan province

作　　者：元正菊 陈红宇 何于雯 孟锦昕 李楠 杨绍昌 王静林 YUAN Zheng-ju;CHEN Hong-yu;HE Yu-wen;MENG Jin-xin;LI Nan;YANG Shao-chang;WANG Jing-lin(Yunnan Animal Science and Veterinary Institute,Yunnan Tropical and Subtropical Animal Virus Disease laboratory,Kunming,Yunnan 650224,China;不详)

机构地区：[1]云南省畜牧兽医科学院,云南省热带亚热带动物病毒重点实验室,云南昆明650224

出　　处：《中国国境卫生检疫杂志》2020年第3期153-156,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：云南省科技厅重点基金项目(2019FA015);国家自然科学基金项目(31660714,31460669);国家重点研发计划项目(2016YFD0500303);云南省科技人才和平台计划项目(2018HB046)。

摘　　要：目的对云南省中缅边境地区采集的蚊虫进行版纳病毒(BAV)的分离鉴定。方法2013年6月在云南省德宏州芒市牛圈采集蚊虫样本,分别接种BHK-21、C6/36、Vero细胞分离病毒,用RT-PCR方法鉴定,并进行序列分析。结果从采集的蚊虫中分离到2株分离物(C23、C36),接种C6/36细胞6 d后产生细胞规律病变,而在BHK21和Vero细胞上不出现细胞病变;PAGE电泳显示2株新分离病毒为12个节段双链RNA病毒,其带型为6-4-2;BAV第12节段特异性引物扩增为阳性,测序获得2株新分离病毒第12节段序列,序列分析显示2株新分离病毒序列与BAV位于相同的进化簇中,核苷酸同源性在92.1%~96.3%之间;与云南省和越南分离的7株BAV遗传进化关系较近,核苷酸同源性在94.5%~96.3%之间,而与我国辽宁、甘肃和山西分离的BAV遗传进化关系较远,核苷酸同源性均低于93.6%。结论从芒市蚊虫中分离的C23和C36病毒为BAV,遗传进化关系与云南省以前分离的毒株较近。Objective To isolate and identify Banna virus(BAV)from mosquitoes collected in China-Myanmar border region of Yunnan province.Methods The mosquito samples were collected from cattle farm in Mangshi,Dehong prefecture,Yunnan in June 2013.The samples were inoculated with BHK-21,C6/36 and Vero cell lines to isolate the virus,using PCR and other methods for identification and sequence analysis.Results Two isolates(C23,C36)were isolated from the collected samples.Regular cell lesions were observed 6 days after inoculation of C6/36 cell line,while no cytopathic effect appeared on BHK-21 and Vero cell line.The results of PAGE gel electrophoresis showed that the two newly isolates possessed 12 segments of double-strand RNA with a banding pattern of 6-4-2.The specific primers of the 12 th segment of BAV were amplified to be positive,and the 12 th segment sequences of the two newly isolated virus obtained.The sequence analysis showed that the sequences of two newly isolates were located in the same evolutionary cluster with BAV strains,and the nucleotide homology was between 92.1%to 96.3%.The genetic and evolutionary relationship between the two newly isolated viruses and 7 strains of BAV isolated from Yunnan province and Vietnam was close,with nucleotide homology between 94.5%to 96.3%,while the genetic and evolutionary relationship between the two strains and 7 strains isolated from Liaoning,Gansu and Shanxi provinces was far,with nucleotide homology lower than 93.6%.Conclusion The C23 and C36 strains isolated from Mangshi mosquito samples were Banna virus,and the genetic and evolutionary relationships were close to those Banna virus strains isolatd in Yunnan province.

关 键 词：版纳病毒 分离鉴定 边境 东南亚十二节段RNA病毒 

分 类 号：R373.3[医药卫生—病原生物学]