蓝舌病病毒7型毒株的分离与分子鉴定

doi:10.11843/j.issn.0366-6964.2017.07.012

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (7): 1281-1287.doi: 10.11843/j.issn.0366-6964.2017.07.012

蓝舌病病毒7型毒株的分离与分子鉴定

吕敏娜^1#, 杨恒^2#, 孙铭飞¹, 李娟¹, 林栩慧¹, 张健騑¹, 廖申权¹, 戚南山¹, 吴彩艳¹, 李华春^2*, 陈琴苓^1*

1. 广东省农业科学院动物卫生研究所广东省兽医公共卫生公共实验室广东省畜禽疫病防治研究重点实验室, 广州 510640;
2. 云南省畜牧兽医科学院, 昆明 650224

收稿日期:2016-11-15 出版日期:2017-07-23 发布日期:2017-07-20
通讯作者: 李华春,E-mail:lihuachun@hotmail.com;陈琴苓,E-mail:544021009@qq.com
作者简介:吕敏娜(1967-),女,广东潮州人,副研究员,硕士,主要从事预防兽医学研究,E-mail:gdczmn@163.com;杨恒(1978-),男,四川西昌人,副研究员,博士,主要从事动物虫媒病毒研究。
基金资助:
公益性行业（农业）科研专项（201303035）；广东省科技计划项目（2015A020209075；2015A020209084）

Isolation and Identification of Bluetongue Virus Serotype 7 Field Isolate

LÜ Min-na^1#, YANG Heng^2#, SUN Ming-fei¹, LI Juan¹, LIN Xu-hui¹, ZHANG Jian-fei¹, LIAO Shen-quan¹, QI Nan-shan¹, WU Cai-yan¹, LI Hua-chun^2*, CHEN Qin-ling^1*

1. Guangdong Open Laboratory of Veterinary Public Health, Guangdong Provincial Key Laboratory of Animal Disease Prevention, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
2. Yunnan Animal Science and Veterinary Institute, Kunming 650224, China

Received:2016-11-15 Online:2017-07-23 Published:2017-07-20
Supported by:

摘要/Abstract

摘要：

为监测广东省蓝舌病流行状况，2014年从广东省某一奶牛场采集血液样品，检测虫媒病病原，对经RT-PCR检测蓝舌病病毒核酸阳性的样品进行病毒分离并鉴定。经鸡胚静脉接种后取肝，研磨后离心取上清转接C6/36细胞和BHK-21细胞，出现CPE，RT-PCR检测蓝舌病病毒的VP7基因并进行序列比对，电镜观察病毒粒子，证实分离到蓝舌病病毒，命名为GD/ST2014。进一步扩增VP2基因，进行系统发育树分析，以及血清中和试验，表明此分离株为血清型7型。这是国内首次报道牛源蓝舌病病毒血清型7型的分离鉴定。

关键词: 蓝舌病病毒, 分离, 鉴定, 血清型, 序列分析

Abstract:

In 2014, cattle blood samples were collected from Guangdong province and vector-borne pathogens contained in them were detected in the present study. Samples which were detected as BTV positive by quantitative reverse transcription polymerase chain reaction (qRT-PCR) were isolated and identified. Embryonated chicken eggs were subsequently inoculated and their livers were harvested to passage on C6/36 and BHK-21 cells, respectively. Extensive cytopathic changes were observed. A subsequent qRT-PCR assay target with VP 7 gene and SEM observation results further confirmed the presence of BTV, and named as GD/ST2014. In addition, VP 2 gene fragment was used for genotyping. And combined with VNT test results, the genotype of the isolate obtained in the present study were identified as BTV-7. This was the first report that bluetongue virus serotype 7 field isolate was isolated from cattle in China.

Key words: bluetongue virus, isolation, identification, serotype, sequence analysis

中图分类号:

S852.659.4

吕敏娜, 杨恒, 孙铭飞, 李娟, 林栩慧, 张健騑, 廖申权, 戚南山, 吴彩艳, 李华春, 陈琴苓. 蓝舌病病毒7型毒株的分离与分子鉴定[J]. 畜牧兽医学报, 2017, 48(7): 1281-1287.

Lü Min-na, YANG Heng, SUN Ming-fei, LI Juan, LIN Xu-hui, ZHANG Jian-fei, LIAO Shen-quan, QI Nan-shan, WU Cai-yan, LI Hua-chun, CHEN Qin-ling. Isolation and Identification of Bluetongue Virus Serotype 7 Field Isolate[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(7): 1281-1287.

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蓝舌病病毒7型毒株的分离与分子鉴定

Isolation and Identification of Bluetongue Virus Serotype 7 Field Isolate

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