中国兽医科学2018，Vol.48Issue(1)：7-12,6.DOI:10.16656/j.issn.1673-4696.2018.0002
猪繁殖与呼吸综合征病毒RT-LAMP可视化检测方法的建立与应用
Establishment and applications of RT-LAMP for the visual detection of porcine reproductive and respiratory syndrome virus
摘要
Abstract
This study was to establish a detection method for porcine reproductive and respiratory syndrome virus.Three sets of specific primers were designed according to the conserved region sequence of porcine reproductive and respiratory syndrome virus(PRRSV) ORF7 gene.An RT-loop-mediated isothermal nucleic acid amplification(RT-LAMP) detection method was established by optimizing the reaction components,such as Bst DNApolymerase,primers,Mg2+dNTPs,SYBR Green Ⅰ and HNB dye concentration.The reaction was carried out at 62℃ for 35 min.The amplification products could be observed as white precipitate,and the ladder strip could be shown by 1.5％agarose gel electrophoresis.The detection limit of this method was 1 × 100.5 TCID50/mL.There were no cross-reactions with CSFV,PRV,PCV 2 and PEDV.The RT-LAMPwas used to detect PRRSV in 110 clinical samples,of which the positive rate was 60.0％.And the coincidence rate with RT-PCR was 85.5％.The method is proved to be simple,sensitive and can be used for rapid detection of PRRSV.关键词
猪繁殖与呼吸综合征病毒/反转录-环介导等温核酸扩增(RT-LAMP)/猪/ORF7基因Key words
porcine reproductive and respiratory syndrome virus(PRRSV)/RT-LAMP/porcine/0RF7 gene分类
农业科技引用本文复制引用
张日腾,张乔亚,姜辰龙,刘宗霞,杨茂春,姜平,白娟..猪繁殖与呼吸综合征病毒RT-LAMP可视化检测方法的建立与应用[J].中国兽医科学,2018,48(1):7-12,6.基金项目
国家自然科学基金项目(31672565,31502082) （31672565,31502082）
国家生猪现代产业体系建设专项(CARS-36) （CARS-36）
