猪嗜淋巴疱疹病毒3型TaqMan荧光定量PCR检测方法的建立及应用  

Development and application of TaqMan real-time PCR for detection of the porcine lymphotropic herpesviruses 3

作　　者：朱事康[1] 佟铁铸[1] 刘星[1] 李春萍[1] 陈燕忠[1] 罗卓军[1] 林志雄[2] 刘中勇[2] 周宇[1] 

机构地区：[1]惠州出入境检验检疫局技术中心,广东惠州516006 [2]广东出入境检验检疫局,广东广州510623

出　　处：《畜牧与兽医》2018年第2期67-71,共5页Animal Husbandry & Veterinary Medicine

基　　金：广东省科技厅项目(2013B020307009)

摘　　要：依据GenBank公布的猪嗜淋巴疱疹病毒3型（porcine lymphotropic herpesviruses 3,PLHV-3）DNA序列,设计特异性引物,优化反应条件,建立荧光定量PCR方法,并对方法的灵敏度、特异性和重复性进行试验。得出方法的线性范围为：8.2×101-8.2×107,灵敏度可达82个拷贝,方法的特异性高,只能检测pMD18-T/PLHV3重组质粒,另外5种对照病原均未检出;方法的重复性好,对不同浓度的pMD18-T/PLHV3重组质粒分别扩增6次,结果重复性好。使用建立的方法检测100份临床样品,同时与普通PCR方法比较,显示荧光定量PCR检出率比普通PCR高,具有较高的灵敏度和准确性。结果表明：建立的荧光定量PCR可用于PLHV-3的临床快速诊断。The porcine lymphotropic herpesviruses（PLHV） have been found to be associated with porcine post-transplant lymphoproliferative disease. PLHV are of high prevalence in pig populations world-wide. Here,two pairs of primers were designed based on the PLHV-3 strains in Gen Bank. A Taq Man-based real-time quantitative PCR（q PCR） assay was developed to detect the specificity of PLHV-3. The results showed that the developed RT-PCR could detect 82 copies of plasmid DNA,and was of higher specificity,sensitivity,accuracy and repeatability which were proved by comparing our q PCR and the conventional PCR in testing 100 clinic samples. The q PCR could be applied to immediate diagnosis of PLHV-3 in clinical cases.

关 键 词：猪嗜淋巴疱疹病毒3型 TAQMAN 荧光定量PCR 

分 类 号：S852.6[农业科学—基础兽医学]