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Rapid detection of highly pathogenic porcine reproductive and respiratory syndrome virus by a fluorescent probe-based isothermal recombinase polymerase amplification assay

Yang, Yang, Qin, Xiaodong, Sun, Yingjun, Chen, Ting, Zhang, Zhidong
Virus genes 2016 v.52 no.6 pp. 883-886
Classical swine fever virus, Foot-and-mouth disease virus, Porcine reproductive and respiratory syndrome virus, RNA, Suid herpesvirus 1, cross reaction, detection limit, fluorescence, quantitative polymerase chain reaction, rapid methods, reverse transcriptase polymerase chain reaction, reverse transcription
A novel fluorescent probe-based real-time reverse transcription recombinase polymerase amplification (real-time RT-RPA) assay was developed for rapid detection of highly pathogenic type 2 porcine reproductive and respiratory syndrome virus (HP-PRRSV). The sensitivity analysis showed that the detection limit of RPA was 70 copies of HP-PRRSV RNA/reaction. The real-time RT-RPA highly specific amplified HP-PRRSV with no cross-reaction with classic PRRSV, classic swine fever virus, pseudorabies virus, and foot-and-mouth disease virus. Assessment with 125 clinical samples showed that the developed real-time RT-RPA assay was well correlated with real-time RT-qPCR assays for detection of HP-PRRSV. These results suggest that the developed real-time RT-RPA assay is suitable for rapid detection of HP-PRRSV.