对虾病毒DNA/RNA的实时荧光定量RT-PCR同步检测方法研究  

Study on a Real-time RT-PCR for Detection of DNA/RNA Viruses of Shrimp

作　　者：沈飚 王忠发[2] 胡兴娟 顾松叶[2] 

机构地区：[1]舟山出入境检验检疫局,浙江舟山316000 [2]舟山市疾病预防控制中心,浙江舟山316021

出　　处：《河南农业科学》2013年第10期146-148,共3页Journal of Henan Agricultural Sciences

摘　　要：为建立同步检测对虾DNA/RNA病毒的实时荧光定量反转录PCR(RT-PCR)方法,提高检验检疫工作效率,探索了一种新的实时荧光定量PCR扩增反应程序。结果显示,应用优化的实时荧光定量RT-PCR检测4种对虾病毒(白斑综合症病毒、传染性皮下及造血器官坏死病毒,桃拉综合征病毒、黄头病毒)DNA/RNA,扩增曲线形态典型,扩增效率理想(E值分别为1.06、1.07、0.92、0.92),标准曲线线性良好(r=1),重复性一致(标准差0.05～0.46,变异系数0.26%～1.62%),灵敏度高,且与实时荧光定量PCR相比差异不显著(平均Ct值误差0.04～0.40,t值0.53～2.50,P>0.05),检测时间约1h。优化的荧光定量RT-PCR可以用于4种对虾病毒DNA/RNA的快速定量检测。To increase the inspection and quarantine efficiency of DNA/RNA viruses of shrimp,a real-time RT-PCR assay was established and optimized for simultaneously detecting four viruses （WSSV,IHHNV,TSV,YHV） of shrimp. The optimized real-time RT-PCR generates typical am- plification curves,ideal amplification efficiencies （E= 1.06,1.07,0.92,0.92）, well linear standard curve （r= 1）, uniform repeatability （Standard deviation distribution between 0.05 and 0.46, vari- ation coefficient distribution at 0.26%- 1.62%）, and high sensitivity, with no significant difference compared with the real-time PCR. The total detection time is about 1 h. The optimized realtime RT-PCR assay can be used for the rapid detection of 4 kinds of shrimp -viruses.

关 键 词：荧光定量RT—PCR 对虾病毒 同步扩增DNA RNA 

分 类 号：S945.4[农业科学—水产养殖]