摘要
为了解我国云南省蓝舌病病毒(BTV)的流行情况和基因分型,本研究在云南师宗、江城、芒市分别设立监控点,通过对哨兵动物定期采血监控其血清学转阳情况,采用"鸡胚-C6/36细胞-BHK-21细胞"接种方式分离病毒,利用qRT-PCR及中和试验鉴定病毒,通过特异性引物对分离株Seg-2及Seg-7序列的ORF区进行RT-PCR扩增与克隆测序。结果显示,从2017年6月师宗县牛的抗凝血中分离到一株BTV,鉴定为BTV-2型,TCID50为10-4.88/50μL。分离株Seg-2及Seg-7序列的系统发生树显示其Seg-2序列和日本以及台湾BTV-2株的Seg-2序列相似度较高,同为BTV-2型,而Seg-7序列和台湾BTV-2株以及日本BTV-21株亲缘关系最近,同为Eastern 2型。本研究分离鉴定了BTV-2型病毒株YN/SZ/22457,并掌握了分离株Seg-2及Seg-7序列的遗传特性,明确了云南省存在BTV-2的流行。本研究为进一步开展BTV-2型的流行病学、感染特性研究奠定了基础。
In order to investigate the prevalence and genetic characteristics of bluetongue virus(BTV)in Yunnan province of China.Three BTV monitoring points were set up at Shizong,Jiangcheng and Mangshi in Yunnan province,blood samples were collected regularly to monitor the serological status of the sentinel animals.Virus isolation was carried out using the"chicken embryo-C6/36 cells-BHK-21 cells"inoculation method,subsequently qRT-PCR and serum neutralization test were used to identify the virus.The Seg-2 and Seg-7 gene segments of the isolate were subjected to RT-PCR amplification and sequencing with specific primers.Results showed that a strain of BTV was isolated from Shizong in June 2017 and identified as serotype BTV-2,the TCID50 of the virus is 10-4.88/50μL.The phylogenetic analysis of the Seg-2 and Seg-7 sequences of YN/SZ/22457 showed that the Seg-2 had highest similarity to Japan and Taiwan BTV-2 strains,while the Seg-7 sequence had the closest relationship with Taiwan BTV-2 strain and Japan BTV-21 strain,both are of"Eastern 2".Here,a BTV-2 strain was isolated in Yunnan province,the genetic characteristics of Seg-2 and Seg-7 sequences were analyzed,confirming the prevalence of BTV-2 in Yunnan province.This study lays a foundation for further research of epidemiology and infection characteristics of Chinese BTV-2.
作者
朱沛
李占鸿
谢佳芮
牛保生
姚萍芬
朱建波
肖雷
李华春
ZHU Pei;LI Zhan-hong;XIE Jia-rui;NIU Bao-sheng;YAO Ping-fen;ZHU Jian-bo;XIAO Lei;LI Hua-chun(Yunnan Tropical and Subtropical Animal Virus Disease Laboratory Yunnan Animal Science and Veterinary Institute,Kunming 650224,China;Shizong Center for Animal Disease Control and Prevention,Shizong 655700,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2020年第5期457-461,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
云南省应用基础研究计划青年项目资助项目(2017FD056)
国家公益性行业项目(农业)专项(201303035)
关键词
BTV2型
分离
鉴定
序列分析
BTV-2
isolation
identification
sequence analysis
作者简介
朱沛(1991-),女,湖南娄底人,硕士,助理研究员,主要从事动物病毒学研究;通信作者:李华春,E-mail:Li_huachun@hotmail.com;通信作者:肖雷,E-mail:xl914204@126.com
