摘要
为同时检测患病大口黑鲈中不同属虹彩病毒感染和带毒情况,针对蛙病毒属和肿大细胞病毒属大口黑鲈虹彩病毒主衣壳蛋白(major capsid protein,MCP)基因的序列,分别设计1对特异性引物Rana-mcp F/R和Mega-mcp F/R,扩增产物片段大小分别为475 bp和262 bp。通过PCR反应体系的优化、反应的特异性和灵敏度试验,建立一种可以同时检测大口黑鲈蛙病毒属和肿大细胞病毒属虹彩病毒感染的双重PCR检测方法,最低DNA检测量分别为6.5 pg和14.5 pg。用此方法,对临床获得的15个大口黑鲈样品进行双重PCR检测和序列测定,获得5个蛙病毒属和1个肿大细胞病毒属虹彩病毒检测阳性结果。本研究中建立的基于MCP基因的大口黑鲈虹彩病毒双重PCR检测方法,可用于养殖大口黑鲈虹彩病毒病快速鉴别诊断和分子流行病学调查。
In order to detect different species of iridescent virus from largemouth bass,two pairs of specific primers,named Rana-mcp and Mega-mcp with predicted products of 475 bp and 262 bp,were designed from sequences of major capsid protein（MCP） gene from Ranavirus and Megalocytivirus,respectively.The reaction conditions of the duplex PCR were optimized,and the specificity and sensitivity were further studied.The limit DNA of the duplex PCR for Ranavirus and Megalocytivirus were 6.5 pg and 14.5 pg,respectively.Using duplex PCR and sequencing methods,15 clinical samples were tested and 5 positive samples from Ranavirus infection and 1 positive sample from Megalocytivirus infection were detected.The developed duplex PCR method based on MCP gene could be used for rapid diagnosis and epidemiology investigation on iridescent virus disease of cultured largemouth bass.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2013年第4期106-110,共5页
Journal of Huazhong Agricultural University
基金
广东省省级财政鱼病防治专项和现代产业技术体系建设专项(CARS-46)
