猪瘟病毒SYBR Green I实时荧光定量PCR检测方法的建立和初步应用  

Development and Application of SYBR Green I Real-Time PCR Technique for Detecting Hog Cholera Virus

作　　者：王荣[1,2] 孔繁德[2] 陈琼[3] 吴德峰[1] 徐淑菲[2] 

机构地区：[1]福建农林大学动物科学学院,福州350002 [2]厦门出入境检验检疫局,厦门361012 [3]厦门市农产品质量安全检验测试中心,厦门361009

出　　处：《经济动物学报》2009年第1期39-42,共4页Journal of Economic Animal

基　　金：厦门出入境检验检疫局科技计划项目(XK09-2003)

摘　　要：根据GenBank已发表的HCV 5’NTR核苷酸序列设计一对特异性引物，扩增位置在整个基因的221～377位，片段长度为167bp。提取由厦门出入境检验检疫局技术中心动物实验室保存的HCV细胞毒的RNA，并反转录成cDNA，以此cDNA为模板建立了SYBR Green I实时定量PCR检测猪瘟病毒的方法，并用该方法检测送检的临床疑似病料。结果表明：本研究建立的HCV-SYBR Green I PCR特异性强，与FMDV、PCV2、PRRSV、PPV、PRV等常见猪病毒病病原没有交叉反应；灵敏度高，可以检测到5．3×10^2的病毒拷贝数；操作简单，耗时短，只需3h即可完成整个试验过程，可初步用于临床检测。According to the 5＇NTR sequences of hog cholera virus（HCV） in GenBank, a pair of special primers were designed, the fragment of amplification should be 167 bp in length. Total RNA was extracted from the cells cultured HCV and then was reversed transcription to cDNA. HCV SYBR Green I PCR method is developed to detect HCV and clinic samples. The result showed that HCV-SYBR Green I PCR had many advantages such as rapidness, simple operation, high sensitivity and specificity, so the method could be applied to clinic detection of HCV.

关 键 词：SYBR Green I HCV 检测 

分 类 号：S852.651[农业科学—基础兽医学]