中国兽医科学2011，Vol.41Issue(10)：1021-1025,5.
猪流感病毒一步法荧光定量RT-PCR诊断方法的建立
Development of one-step real-time quantitative RT-PCR approach for the detection of swine influenza virus
摘要
Abstract
In order to develop a rapid and high-performance assay method for epidemiological study and clinical diagnosis of swine influenza virus,a one-step real-time quantitative RT-PCR method based on a TaqMan probe was developed.Conserved regions in the M gene of swine influenza viruses served as targets for primers and TaqMan probe.Concentrations of primers and probe were optimized to improve the sensitivity and specificity of reactions.A series of dilutions of recombinant plasmid were prepared and used to generate standard curve with an efficiency value of 2.014 and a error value of 0.005.Results showed that the threshold of 10 copies of target molecules can be detected,which revealed high sensitivity of the method.Clinical diagnosis showed that this approach was more sensitive than normal RT-PCR and virus isolation methods.Besides,it was negative for another six popular viruses in swine,which showed high specify of the approach,Moreover,it was easy to be operated and was time-saving.In conclusion,the method established in the present study can be used for rapid detection and monitoring in epidemiology study of SIV.关键词
猪流感病毒/一步法荧光定量RT-PCR/TaqMan探针Key words
swine influenza virus/one-step real-time quantitative RT-PCR/TaqMan probe分类
农业科技引用本文复制引用
徐敏,李志强,黄元,陈晶,陈少勤,张健騑,赵亚华,宣华,向华..猪流感病毒一步法荧光定量RT-PCR诊断方法的建立[J].中国兽医科学,2011,41(10):1021-1025,5.基金项目
广东省农业科学院院长基金项目（20090203) （）
广东省科技厅项目（2009B020600001、2009A020101006、2006B20801002、2006A20301006、2006B21101004) （）
东莞市科技局项目（200910810130) （）
