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非洲猪瘟病毒锁核酸探针荧光定量PCR检测方法的建立
引用本文:李艳,郭怡德,勾红潮,卞志标,孙铭飞,蔡汝健,宋帅,蒋智勇,楚品品,徐民生,杨东霞,李春玲. 非洲猪瘟病毒锁核酸探针荧光定量PCR检测方法的建立[J]. 中国兽医学报, 2021, 0(2): 208-212,230
作者姓名:李艳  郭怡德  勾红潮  卞志标  孙铭飞  蔡汝健  宋帅  蒋智勇  楚品品  徐民生  杨东霞  李春玲
作者单位:1.广东省农业科学院动物卫生研究所;2.广东省畜禽疫病防治研究重点实验室;3.农业农村部兽用药物与诊断技术广东科学观测实验站
基金项目:广东省自然科学基金资助项目(2020A1515010475);广东省重点领域研发计划资助项目(2019B020211005,2019B020217002);国家重点研发计划资助项目(2016YFD0500709)。
摘    要:根据非洲猪瘟病毒(African swine fever virus,ASFV)P72基因核苷酸序列设计特异性引物和锁核酸(locked nucleic acid,LNA)-TaqMan探针,建立了基于P72基因的LNA-TaqMan探针的ASFV荧光定量PCR方法.结果显示,所建立的LNA-TaqMan探针荧光定量P...

关 键 词:非洲猪瘟病毒  P72基因  LNA-TaqMan探针荧光定量PCR

Establishment of LNA-TaqMan probe real-time PCR for detection of African swine fevervirus
LI Yan,GUO Yide,GOU Hongchao,BIAN Zhibiao,SUN Mingfei,CAI Ru-jian,SONG Shuai,JIANG Zhiyong,CHU Pinpin,XU Minsheng,YANG Dongxia,LI Chunling. Establishment of LNA-TaqMan probe real-time PCR for detection of African swine fevervirus[J]. Chinese Journal of Veterinary Science, 2021, 0(2): 208-212,230
Authors:LI Yan  GUO Yide  GOU Hongchao  BIAN Zhibiao  SUN Mingfei  CAI Ru-jian  SONG Shuai  JIANG Zhiyong  CHU Pinpin  XU Minsheng  YANG Dongxia  LI Chunling
Affiliation:(Institute of Animal Health,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;Key Laboratory of Livestock Disease Prevention of Guangdong Province,Guangzhou 510640,China;Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province,Ministry of Agriculture,Guangzhou 510640,China)
Abstract:In this study,LNA-TaqMan probe-based real-time PCR was established using specific primers and probes according to the P72 gene of African swine fever virus(ASFV).The results showed that the LNA-TaqMan probe-based real-time PCR had higher sensitivity,and the minimum detection limit was 3.9 copies/μL.The LNA-TaqMan probe-based real-time PCR was no cross-reaction with various pathogens such as swine fever virus,porcine reproductive and respiratory syndrome virus,and porcine circovirus type 2.The intra-and inter-assay coefficient of variation was less than 1%,with good repeatability.The results of 56 clinical samples were consistent with the results of qPCR recommended by OIE,and the coincidence rate was 100%.The LNA-TaqMan probe-based real-time PCR established in this study had ideal sensitivity,specificity,and repeatability,which provides a new technology choice for the detection of ASFV.
Keywords:ASFV  P72 gene  LNA-TaqMan real-time PCR
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