Pyrococcus furiosus Argonaute-mediated porcine epidemic diarrhea virus detection

Abstract

Porcine epidemic diarrhea virus (PEDV), an enteric coronavirus, induces severe vomiting and acute watery diarrhea in unweaned piglets. The pig industry has suffered tremendous financial losses due to the high mortality rate of piglets caused by PEDV. Consequently, a simple and rapid on-site diagnostic technology is crucial for preventing and controlling PEDV. This study established a detection method for PEDV using recombinase-aided amplification (RAA) and Pyrococcus furiosus Argonaute (PfAgo), which can detect 100 copies of PEDV without cross-reactivity with other pathogens. The entire reaction of RAA and PfAgo to detect PEDV does not require sophisticated instruments, and the reaction results can be observed with the naked eye. Overall, this integrated RAA-PfAgo cleavage assay is a practical tool for accurately and quickly detecting PEDV. Key points • PfAgo has the potential to serve as a viable molecular diagnostic tool for the detection and diagnosis of viral genomes • The RAA-PfAgo detection technique has a remarkable level of sensitivity and specificity • The RAA-PfAgo detection system can identify PEDV without needing advanced equipment

Pyrococcus furiosus Argonaute Based Detection Assays for Porcine Deltacoronavirus

“…Recombinase-aided amplification (RAA) technology and reverse transcription recombinase-aided amplification (RT-RAA) are to obtain recombinase from bacteria or fungi, and realize DNA or RNA amplification in vitro by cooperating with other enzymes. , In this work, RAA and RT-RAA were employed to amplify the nucleic acid before attempting PfAgo testing, since nucleic acid amplification is crucial for sensitivity in nucleic acid testing. Although RAA-PfAgo assay methods have been previously established for detecting virus, − in this study, placing the RT-RAA reaction and PfAgo in the same tube undoubtedly reduced the procedure steps as well as aerosol contamination. In this article, the terms RAA-PfAgo, RT-RAA-PfAgo, and RT-RAA-PfAgo-LFD were used to refer to these three PDCoV detection methods.…”