Pyrococcus furiosus Argonaute Based Detection Assays for Porcine Deltacoronavirus

ACS Synth Biol. 2024 Apr 19;13(4):1323-1331. doi: 10.1021/acssynbio.4c00045. Epub 2024 Apr 3.

Abstract

Porcine deltacoronavirus (PDCoV) is a major cause of diarrhea and diarrhea-related deaths among piglets and results in massive losses to the overall porcine industry. The clinical manifestations of porcine diarrhea brought on by the porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), and PDCoV are oddly similar to each other. Hence, the identification of different pathogens through molecular diagnosis and serological techniques is crucial. Three novel detection methods for identifying PDCoV have been developed utilizing recombinase-aided amplification (RAA) or reverse transcription recombinase-aided amplification (RT-RAA) in conjunction with Pyrococcus furiosus Argonaute (PfAgo): RAA-PfAgo, one-pot RT-RAA-PfAgo, and one-pot RT-RAA-PfAgo-LFD. The indicated approaches have a detection limit of around 60 copies/μL of PDCoV and do not cross-react with other viruses including PEDV, TGEV, RVA, PRV, PCV2, or PCV3. The applicability of one-pot RT-RAA-PfAgo and one-pot RT-RAA-PfAgo-LFD were examined using clinical samples and showed a positive rate comparable to the qPCR method. These techniques offer cutting-edge technical assistance for identifying, stopping, and managing PDCoV.

Keywords: PfAgo; detection; porcine deltacoronavirus.

MeSH terms

  • Animals
  • Coronavirus Infections* / diagnosis
  • Coronavirus Infections* / veterinary
  • Deltacoronavirus*
  • Diarrhea / diagnosis
  • Porcine epidemic diarrhea virus*
  • Pyrococcus furiosus* / genetics
  • Recombinases
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases* / diagnosis
  • Swine Diseases* / epidemiology

Substances

  • Recombinases

Supplementary concepts

  • Porcine coronavirus HKU15