Real-time Fluorescence Reverse-transcription Loop-mediated Isothermal Amplification for Detection of Porcine Epidemic Diarrhea Virus

摘要

cqvip:Porcine epidemic diarrhea,a highly contagious enteric infectious disease caused by the porcine epidemic diarrhea virus( PEDV) with symptoms of vomit,diarrhea,loss of appetite of suckling pig,has led to serious economic loss to the global swine industry. In this study,a real-time fluorescence reverse transcription loop-mediated isothermal amplification( RT-LAMP) assay was developed to detect PEDV RNA. The real-time fluorescence RT-LAMP assay was performed at62 ℃ for 60 min,using a simple and portable device,the ESE-Quant Tube Scanner. The detection limit of RNA was 2. 9 × 10~6 copies/μl,10 times as sensitive as RT-PCR,and the detection was specific only to PEDV. Application of this method to clinical samples yielded a positivity rate of 93%,which was higher than that of RT-PCR. This technique saves time and is efficient,and is thus expected to be useful for the diagnosis of PEDV infection in the field.