中国畜牧兽医2017，Vol.44Issue(11)：3320-3326,7.DOI:10.16431/j.cnki.1671-7236.2017.11.030
猪细小病毒重组酶聚合酶扩增快速检测方法的建立
Development of the Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Parvovirus
摘要
Abstract
This study was aimed to develop a simple and rapid method for the porcine parvovirus (PPV) with recombinase polymerase amplification (RPA),which could be a novel and reliable tool for the control and detect of PPV.The RPA was developed using specific primers for the conserved region of PPV VP2 gene.The reaction time was optimized,the RPA reaction could amplify the PPV,and was performed successfully at 38 ℃C for 30 min in a water bath.The results showed that the detection limit of RPA was 102 copies of plasmid DNA,which was the same as the Real-time quantitative PCR applied in this study,and 100 times more sensitive than conventional PCR.For the clinical samples from the suspected PPV-infected pigs,the positive detection ratio was 82.6％ for RPA,which was lower than that of Real-time quantitative PCR (86.9％),but was much higher than conventional PCR (66.7％).The PPV RPA assay developed in the study was simple,rapid and reliable,and was suitable for rapid detection of PPV.关键词
猪细小病毒(PPV)/等温扩增/聚合酶重组酶扩增/快速检测Key words
porcine parvovirus (PPV)/isothermal amplification/recombinase polymerase amplification (RPA)/rapid detection分类
农业科技引用本文复制引用
刘立兵,王建昌,经美,王金凤,袁万哲..猪细小病毒重组酶聚合酶扩增快速检测方法的建立[J].中国畜牧兽医,2017,44(11):3320-3326,7.基金项目
河北省自然科学基金青年项目(C2017325001) （C2017325001）
猪主要繁殖障碍性疫病防控关键技术研究(16226604D) （16226604D）
