家畜生态学报2011，Vol.32Issue(2)：50-54,5.
猪繁殖与呼吸综合征病毒LAMP检测方法的建立
Development of a LAMP Assay for Detection of Porcine Reproductive and Respiratory Syndrome Virus
摘要
Abstract
Four pairs of primers were designed based on conserved N protein gene of porcine reproductive and respiratory syndrome virus (PRRSV) American type in this study. Through optimization of various reaction conditions, a loop-mediated isothermal amplification (LAMP) assay was established for the detection of PRRSV. It was shown that the LAMP assay had a good specifity and achieved theoretically a sensitivity of 1 × 101 copies, whereas routine PCR reached 1 × 105 copies. Without the help of expensive machines, the detection result of LAMP could be observed by adding 1 μL orange-red nucleic acid dyes.The positive samples showed green color, while the negative ones kept orange-red color. In summary, the constructed LAMP is a rapid, intuitive and accurate method, which is especially suitable for the detection of PRRSV in the pig farms.关键词
猪繁殖与呼吸综合征病毒/LAMP/核酸染料/现地检测Key words
porcine reproductive and respiratory syndrome virus/ LAMP/ nucleic acid dye/ clinical detection分类
农业科技引用本文复制引用
尹斐斐,时建立,李俊,周顺,王金宝,吴家强,刘少宁,李坤,杜以军,孙文博,丛晓燕,王可,张玉玉..猪繁殖与呼吸综合征病毒LAMP检测方法的建立[J].家畜生态学报,2011,32(2):50-54,5.基金项目
农业科技成果转化资金项目(2010GB2C600259) （2010GB2C600259）
农业科技跨越计划(2009农跨18) （2009农跨18）
山东省自主创新成果转化重大专项(2010ZHZXlA0417) （2010ZHZXlA0417）
山东省农业重大应用技术创新课题(鲁财农便[2010]5号) （鲁财农便[2010]5号）
