Rapid and specific detection of porcine parvovirus by isothermal recombinase polymerase amplification assays.
Porcine parvovirus (PPV) is a major cause of swine reproductive failure and reported in many countries worldwide. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PPV real-time RPA assay) and a lateral flow dipstick (PPV RPA LFD assay) were developed targeting PPV NS1 gene. The detection limit of PPV real-time RPA assay was 300 copies per reaction within 9 min at 38 °C, while the RPA LFD assay has a detection limit of 400 copies per reaction in less than 20 min at 38 °C. In both assays, there were no cross-reactions with porcine circovirus type 2, pseudorabies virus, porcine reproductive and respiratory syndrome virus, classical swine fever virus, and foot-and-mouth disease virus. Based on a total of 128 clinical samples examined, the sensitivity and the specificity of the developed RPA assays for identification of PPV was 94.4% and 100%, respectively, when compared to real-time (qPCR) assay. Therefore, the RPA assay provides a rapid, sensitive and specific alternative for PPV detection.
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Yang
Y
Yang
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. Electronic address: qinxiaodong@caas.cn.
Xiaodong
X
Qin
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Wei
W
Zhang
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. Electronic address: liyanmin@caas.cn.
Yanmin
Y
Li
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Zhidong
Z
Zhang
D000818:Animals; D028323:Parvovirus, Porcine / Q000235:genetics* / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D045522:Recombinases / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013552:Swine; D013696:Temperature
D045522:Recombinases
PPV; RPA LFD assay; Real-time RPA assay; Recombinase polymerase amplification assay