中国动物传染病学报Issue(5)：10-15,6.
猪细环病毒k2型SYBR GreenⅠ实时荧光定量PCR方法的建立
DEVELOPMENT OF A SYBR GREENⅠ QUANTITATIVE REAL-TIME PCR METHOD FOR PORCINE TORQUE TENO SUS VIRUS K2
摘要
Abstract
In order to rapidly and quantitatively detect Porcine Torque teno sus virus k2 (PTTSuVk2) , a SYBR GreenⅠ-based quantitative real time PCR assay was developed using specifi cally paired primers targeting to the ORF2 gene of PTTSuVk2. The PCR assay was linear in the range of 1.92×102~1.92×107 copies per microliter. A series of experiments were carried out to assess the sensitivity, specifi city and reproducibility of the method, followed by the intra-assay and inter-assay CVs for CT values obtained with the standard plasmids. The melting curve analysis using SYBR GreenⅠ dye showed one specifi c peak with a melting temperature (Tm) at (83.83±0.08)℃ without primer-dimers peak. The intra-assay CVs were equal or less than 1.69 % and the inter-assay CVs were equal or less than 2.19 %. No cross-reaction occurred with nucleic acids extracted from Porcine circovirus (type 1 and type 2), Porcine parvovirus,Porcine bovavirus type 5 and Porcine torque teno virus 1a and 1b. The results demonstrated that the assay was specifi c and reproducible, suggesting the quantitative PCR might be used for rapid laboratory diagnosis and epidemiological investigation for PTTSuVk2.关键词
猪细环病毒k2型/ORF2基因/SYBR GreenⅠ/实时荧光定量PCR方法Key words
Porcine Torque teno sus virus k2/ORF2 gene/SYBR Green I/real time PCR分类
农业科技引用本文复制引用
陈如敬,黄秋宇,修金生,吴学敏,严山,车勇良,周伦江..猪细环病毒k2型SYBR GreenⅠ实时荧光定量PCR方法的建立[J].中国动物传染病学报,2016,(5):10-15,6.基金项目
福建省属公益类科研专项 （）
