基于便携式荧光定量PCR仪的兔病毒性出血症病毒现场检测方法的建立及应用  

Establishment and Application of a Live Detection Method of Rabbit Hemorrhagic Disease Virus Based on Portable Fluorescence Quantitative PCR Instrument

作　　者：李天芝 于新友 LI Tianzhi;YU Xinyou(Shandong Lvdu Bio-Industry Co.,Ltd.,Binzhou 256600,Shandong China)

机构地区：[1]山东绿都生物科技有限公司

出　　处：《饲料博览》2019年第12期1-4,共4页Feed Review

基　　金：山东省现代农业产业技术体系特种经济动物创新团队项目（SDAIT-21-15）

摘　　要：为建立兔病毒性出血症病毒免疫荧光PCR检测方法,参照GenBank中兔病毒性出血症病毒基因组序列,设计特异引物和TaqMan探针,通过优化反应条件,建立了能检测兔病毒性出血症病毒的荧光PCR方法,并验证该方法的特异性、敏感性、重复性。结果表明,该方法检测兔病毒性出血症病毒灵敏度可达1.38LD50·100μL-1,该法对兔巴氏杆菌、兔大肠杆菌、兔沙门氏菌、兔水疱性口炎病毒(RVSTV)和兔轮状病毒(RRV)的检测结果均为阴性。试验建立的TaqMan荧光定量PCR检测方法可对兔病毒性出血症进行现场快速鉴别诊断,为兔病毒性出血症的诊断及防控工作奠定了基础。To establish a fluorescent PCR method for detection of Rabbit viral hemorrhagic disease(RVHD).In this study,specific primers and TaqMan probes were designed according to the genomic sequence of Rabbit viral hemorrhagic disease virus in GenBank.By optimizing the reaction conditions,a fluorescent PCR method for detecting rabbit viral hemorrhagic disease virus was established,and the specificity,sensitivity and repeatability of the method were verified.The results showed that the sensitivity of this method for detecting Rabbit viral hemorrhagic disease virus(RVSTV) was up to 1.38 LD50·100 μL-1.The results of this method for detecting rabbit pasteurella,rabbit Escherichia coli,rabbit salmonella,rabbit bullous stomatitis virus(RVSTV) and rabbit rotavirus(RRV) were negative.The TaqMan fluorescent quantitative PCR method established in this experiment can rapidly differentiate and diagnose rabbit viral hemorrhage in the field,which lays a foundation for the needle diagnosis and prevention and control of rabbit viral hemorrhage.

关 键 词：兔病毒性出血症病毒 TagMan 荧光PCR 

分 类 号：S852.65[农业科学—基础兽医学]