摘要
参照GenBank已收录的大鲵蛙病毒MCP基因序列设计特异性引物,建立基于大鲵蛙病毒特异性引物的巢式PCR检测体系。本试验利用大鲵蛙病毒CGSV主要核衣壳蛋白MCP基因保守区,设计内引物和外引物并制备了重组质粒pMD19-T作为阳性对照标准品。灵敏度试验结果显示,巢氏PCR的敏感性较普通PCR的敏感性高1 000倍;而且与草鱼呼肠孤病毒(Grass carp reovirus,GCRV)、鲫鱼疱疹病毒2型(Goldfish herpes virus,CyHV-2)、云斑尖塘鳢虹彩病毒(Marbled sleepy goby iridovirus,MSGIV)、迟钝爱德华菌(Edwardsiella tarda)、嗜水气单胞菌(Aeromonas hydrophila)、维氏气单胞菌(Aeromonas veronii)等无交叉反应。该体系具有简便、快捷、特异性高、低成本等特点,为大鲵蛙病毒检测提供了一项重要的技术手段。
A nested PCR were developed as an efficient method to detect Chinese giant salamander Ranavirus. Two pairs of primers were designed based on the sequence of major capsid proteins （MCP） gene of CGSV. A standard recombinant plasmid pMD19-T was reconstructed for the nes- ted PCR assay. The sensitivity of Nested PCR was 1 000 times more than the conventional PCR. The nested PCR was of high specificity without any cross-reactions with DNA from Grass carp reovirus （GCRV）, Goldfish herpes virus （CyHV-2）, Marbled sleepy goby iridovirus （MSGIV）, Edzvardsiella tarda ,Aeromonashydrophila ,Aeromonas veronii. The nested PCR assay is a signif- icant technology for the detection of ranavirus with its sensitivity and specificity.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2014年第3期406-410,共5页
Chinese Journal of Veterinary Science
基金
四川省科技支撑计划资助项目(2011N20071)
教育部<长江学者和创新团队发展计划>创新团队资助项目(IRTO848)
关键词
大鲵蛙病毒
巢氏PCR
大鲵
检测
~ Chinese giant salamander ranavirus
nested PCR ~ Andrias davidianus Blanchard
detect
