Development of an isothermal recombinase polymerase amplification assay for rapid detection of pseudorabies virus.
Recombinase polymerase amplification assays using real-time fluorescent detection (real-time RPA assay) and lateral flow dipstick (RPA LFD assay) were developed targeting the gD gene of pseudorabies virus (PRV). Both assays were performed at 39 °C within 20 min. The sensitivity of the real-time RPA assay and the RPA LFD assay was 100 copies per reaction and 160 copies per reaction, respectively. Both assays did not detect DNAs from other virus or PRV negative samples. Therefore, the developed RPA assays provide a rapid, simple, sensitive and specific alternative tool for detection of PRV.
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.
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.
Zhiyong
Z
Li
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou 730046, Gansu, China.
Shuaijun
S
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. Electronic address: zhangzhidong@caas.cn.
Zhidong
Z
Zhang
D000818:Animals; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D011557:Pseudorabies / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D045522:Recombinases / Q000235:genetics*; D013552:Swine / Q000821:virology
D045522:Recombinases
PRV; RPA LFD assay; Real-time RPA assay; Recombinase polymerase amplification