中国畜牧兽医2017，Vol.44Issue(12)：3434-3439,6.DOI:10.16431/j.cnki.1671-7236.2017.12.006
猪流行性腹泻病毒实时荧光RPA等温检测方法的建立
Establishment of Real-time Recombinase Polymerase Amplification for Detection of Porcine Epidemic Diarrhoea Virus
摘要
Abstract
To develop a precise and rapid diagnosis method for detecting porcine epidemic diarrhoea virus (PEDV),a series of recombinase polymerase amplification (RPA) primers and exoprobes were established based on the highly conserved M gene of PEDV.Then a Real-time RPA assay was developed to detect PEDV using pUC57 plasmid carrying M gene fragment of PEDV as template,and the membrane or nucleotide capsid proteins from TGEV,PRRSV,PCV2 and CSFV were utilized as control.Then the sensitivity and specificity of this Real-time RPA assay was evaluated.The results showed that the Real-time reaction could detect PEDV specifically at 39 ℃ within 20 min with the detection limit of 10 copies/μL of plasmid DNA,and there was no crossreaction with other control viral pathogens.Besides,the established Real-time PRA method could successfully detecte the PEDV M gene in the plasma and plasma protein power.The Real-time established in this study was simple,rapid and sensitive,which could be a novel and reliable method for diagnosing and control of PED.关键词
猪流行性腹泻病毒/分子检测/重组酶聚合酶扩增(RPA)/等温扩增Key words
porcine epidemic diarrhoea virus (PEDV)/molecular diagnosis/recombinase polymerase amplification (RPA)/isothermal amplification分类
农业科技引用本文复制引用
吕继洲,范燕茹,冯春燕,袁向芬,吴绍强..猪流行性腹泻病毒实时荧光RPA等温检测方法的建立[J].中国畜牧兽医,2017,44(12):3434-3439,6.基金项目
国家质检总局科技计划项目(2015IK309) （2015IK309）
国家科技支撑计划课题(2013BAD12B01) （2013BAD12B01）
