中国畜牧兽医2017，Vol.44Issue(6)：1630-1636,7.DOI:10.16431/j.cnki.1671-7236.2017.06.008
猪繁殖与呼吸综合征病毒RT-LAMP快速检测方法的建立
Establishment of RT-LAMP for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus
摘要
Abstract
In order to diagnose porcine reproductive and respiratory syndrome virus (PRRSV) early, a rapid, sensitive, simple reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was established.Four primers targeting the ORF5 gene of PRRSV were designed.Reverse transcription and amplification of the viral cDNA using Bst DNA polymerase was optimal at a constant temperature of 65 ℃.The output of the RT-LAMP assay was visualized using 1.0% agarose gel electrophoresis and color change after the addition of the SYBR Green Ⅰ dye.The assay was also specific for PRRSV and did not cross react with classical swine fever virus (CSFV), porcine circovirus virus type 2 (PCV2), porcine parvovirus (PPV), porcine pseudorabies virus (PRV).The RT-LAMP method was approximately 100-fold more sensitive than RT-PCR for PRRSV detection.The clinical samples (n=10) were identified both by RT-PCR and RT-LAMP, and the coincidece rate was 100%.Thus, the novel RT-LAMP assay was a rapid, simple, sensitive, specific test for PRRSV, and it could potentially be applied in clinical settings.关键词
逆转录环介导等温扩增(RT-LAMP)/猪繁殖与呼吸综合征病毒(PRRSV)/快速检测Key words
reverse transcription loop-mediated isothermal amplification (RT-LAMP)/porcine reproductive and respiratory syndrome virus (PRRSV)/rapid detection分类
农业科技引用本文复制引用
陈希文,汪谦,尹苗,李莲,罗文涛,杨凤,郭爱伟,周杰珑,王雄清..猪繁殖与呼吸综合征病毒RT-LAMP快速检测方法的建立[J].中国畜牧兽医,2017,44(6):1630-1636,7.基金项目
四川省科技厅项目(2013JY0127) （2013JY0127）
四川省重点实验室项目(ESP1505) （ESP1505）
四川省教育厅项目(16ZA0321) （16ZA0321）
