中国畜牧兽医2017，Vol.44Issue(2)：344-349,6.DOI:10.16431/j.cnki.1671-7236.2017.02.006
猪流行性腹泻病毒环介导等温扩增检测方法的建立及应用
Establishment and Application of Loop-mediated Isothermal Amplification for Rapid Detection of Porcine Epidemic Diarrhea Virus
摘要
Abstract
This study was aimed to establish a loop-mediated isothermal amplification (LAMP) assay for detection of porcine epidemic diarrhea virus (PEDV),and provide a simple,sensitive,accurate and reliable tool for diagnosis of PEDV.The conservative PEDV N gene (GenBank accession number:KT799997) of PEDV was selected as a target to design six specific primers.The reaction system and temperature of LAMP were optimized,and the LAMP method for specific amplification of PEDV was established.Results showed that the PEDV LAMP detection method was established successfully,and it could detect PEDV specifically at 60 ℃ for 60 min,and the detection limit was 91 copies/μL,which was one hundred-fold higher than conventional RT-PCR method.75 clinical samples were detected by LAMP and PCR,respectively,the coincidence of LAMP and PCR was about 97.3％.All the data suggested that the LAMP assay had strong specificity,high sensitivity,simple operation,low equipment requirement,and was suitable for rapid detection of PEDV clinical samples.关键词
猪流行性腹泻病毒/环介导等温扩增技术/检测Key words
porcine epidemic diarrhea virus (PEDV)/loop-mediated isothermal amplification (LAMP)/detection分类
农业科技引用本文复制引用
罗亚坤,梁琳,王静,刘存,刘琪,刘畅,蔺文成,崔尚金..猪流行性腹泻病毒环介导等温扩增检测方法的建立及应用[J].中国畜牧兽医,2017,44(2):344-349,6.基金项目
中国农业科学院创新工程项目(ASTIP-IAS16) （ASTIP-IAS16）
