摘要
通过对GenBank中注册的CSFV序列进行比对分析,针对CSFV Npro/C基因中的保守区域,设计合成了一套RT-LAMP引物,应用含有Npro/C基因的阳性质粒株对RT-LAMP引物进行验证。在成功扩增出特异片段的基础上,用CSFV RNA优化RT-LAMP反应体系和条件,建立了CSFV的可视化RT-LAMP快速检测方法。对建立的方法进行特异性和敏感性试验,结果显示,所建立的方法能够特异地检测出CSFV RNA,检测PRRSV、FMDV和JEV RNA均为阴性,并且可检测低至0.1pg/μL浓度的CSFV RNA。
A suit of primers were designed according to a part of Npro/C genes of CSFV from GenBank.The specific fragments were amplified from positive plasmid pMD-CSFV using LAMP method.The RT-LAMP reaction system and condition were optimized with CSFV RNA,so a visualized RT-LAMP method was established.The results showed that CSFV RNA could be detected specifically and the RNA of PRRSV,FMDV and JEV all were detected negative.As low as 0.1pg/μL CSFV RNA could be detected.
出处
《广东畜牧兽医科技》
2011年第2期26-28,32,共4页
Guangdong Journal of Animal and Veterinary Science
基金
国家自然科学基金项目(30771611
31072137)
教育部"长江学者和创新团队发展计划"创新团队项目(TRT0723)
广东省自然科学基金创新团队项目(5200638)
