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目的对中国-老挝边境口岸地区云南省勐腊县和老挝勐新县采集的蚊虫标本进行版纳病毒检测。方法2009年8月在勐腊县、勐新县人房和畜圈采集蚊虫标本,从蚊虫研磨液中提取RNA,用RT-PCR方法扩增版纳病毒第3、5、7、9、l1、12节段序列,聚丙烯酰胺凝胶电泳检测核酸扩增阳性标本的基因组带型,利用MEGA 4软件进行系统进化分析。结果共检测蚊虫标本80批,其中3批老挝标本扩增到版纳病毒第3、7、11节段核酸序列,聚丙烯酰胺凝胶电泳显示3个阳性分离物的基因组均为12条带双链RNA,系统进化分析显示3个老挝版纳病毒分离物位于2个进化分支,其中1株与越南分离株关系最近。结论首次从老挝蚊虫标本中检测到版纳病毒,老挝分离株与中国、越南分离株之间发生了基因重配。
Objective To detect the specimens of mosquitoes collected from Mengla County, Yunnan Province and Mengxin County, Laos, on the border between Laos and China. Methods In August 2009, samples of mosquitoes were collected from human houses and livestock pens in Mengla County and Mengxin County. RNA was extracted from the mosquito larvae and amplified by PCR-RT-PCR for the third, fifth, seventh, ninth, eleventh, The genomic bands of the positive samples were detected by polyacrylamide gel electrophoresis. The phylogenetic analysis was carried out by MEGA 4 software. Results A total of 80 batches of mosquitoes were detected. Among them, 3 batches of Laos samples were amplified to the 3rd, 7th and 11th nucleotides. The polyacrylamide gel electrophoresis showed that the genomes of the 3 positive isolates were all 12 - RNA, Phylogenetic analysis showed that three Lao Banna virus isolates were located in two evolutionary branches, of which one was the closest to Vietnam. Conclusions The first detection of the BAV virus from Laos mosquitoes and the reassignment between Laos and China and Vietnam isolates.