河南农业科学Issue(6)：128-131,144,5.
猪伪狂犬病毒野毒株 SYBR Green Ⅰ实时荧光定量 PCR 诊断试剂盒的研制
Development of SYBR Green Ⅰ Real-time Quantitative PCR Kit for Detection of PRV Wild-type Strains
摘要
Abstract
According to the gene sequences of gE gene of PRV in GenBank ,one pairs of specific primer was designed for amplifying the specific fragments of gE gene .Then gE gene of PRV amplified by PCR was cloned into pMD18-T vector and it was used as positive standard .After optimization of annealing temperature and primers concentrations ,a SYBR GreenⅠ real-time quantitative PCR kit was developed for detection of PRV wild-type strains .The melting curve analysis using SYBR Green Ⅰdye showed one specific peak ,and primer-dimers peak was not observed .No fragment was amplified from PRV strains depleted of gE gene ,PPV ,PCV-2 ,Escherichia coli ,CSFV ,PRRSV by the SYBR Green Ⅰ real-time quantitative PCR .The PCR kit was highly sensitive in 2.3 × 101 copies/μL DNA .The results revealed that the PCR kit was sensitive ,specific and it could be used to detect PRV wild-type strains in clinical samples .关键词
猪伪狂犬病毒/野毒株/gE基因/荧光定量PCR/诊断试剂盒Key words
PRV/wild-type strains/gE/real-time quantitative PCR/diagnostic kit分类
农业科技引用本文复制引用
余波,周思旋,谭诗文,徐景峨,史开志,杨莉..猪伪狂犬病毒野毒株 SYBR Green Ⅰ实时荧光定量 PCR 诊断试剂盒的研制[J].河南农业科学,2014,(6):128-131,144,5.基金项目
贵州省科技厅农业攻关项目（［2010］3085）；贵州省畜禽健康养殖技术创新能力建设项目 （）
