登录GenBank下载猪圆环病毒2型(Porcine circovirus type2,PCV-2)的全基因组序列,根据其保守区域,设计1对特异性引物,采用SYBR Green Ⅰ随机结合渗入法,建立该模式的实时定量PCR检测方法,构建了检测PCV-2型的标准DNA模版,循环阈值(Ct)与标准DNA模板在1.0×102～1.0×107拷贝·μL-1浓度范围内呈良好的线性关系,相关系数为0.999。该方法用于猪圆环病毒2型的检测具有很高的特异性,其敏感性与常规PCR相比可以提高100倍,可以用于猪圆环病毒2型的快速检测。
To establish a TaqMan-based real-time RT-PCR method for detection of canine distemper virus,the pair of primers and the TaqMan-based probe were designed according to the M protein genes of canine distemper virus strains.The reactive conditions were optimized to improve the sensitivity and specificity of the method.The specificity and sensitivity of the method were high.Canine distemper virus in the thirty-eight samples had been tested by the TaqMan-based real-time RT-PCR,the common RT-PCR and electron microscopy.The sensitivity of the TaqMan-based real-time RT-PCR was higher than the common RT-PCR and electron microscopy.This suggests that the method may be used in clinical diagnosis,epizootic study and laboratory research.
