Development of a recombinase polymerase amplification fluorescence assay to detect feline coronavirus

Mol Cell Probes. 2020 Dec:54:101669. doi: 10.1016/j.mcp.2020.101669. Epub 2020 Oct 22.

Abstract

Feline coronavirus (FCoV) is classified into two pathotypes: the avirulent feline enteric coronavirus (FECV), and the virulent feline infectious peritonitis virus (FIPV). Rapid pathogen detection, which is efficient and convenient, is the best approach for early confirmatory diagnosis. In this study, we first developed and evaluated a rapid recombinase polymerase amplification (RPA) detection method for FCoV that can detect FCoV within 15 min at 39 °C. The detection limit of that assay was 233 copies/μL DNA molecules per reaction. The specificity was high: it did not cross-react with canine distemper virus (CDV), canine coronavirus (CCoV), canine adenovirus (CAV), feline calicivirus (FCV), feline herpesvirus (FHV), or feline parvovirus (FPV). This assay was evaluated using 42 clinical samples (30 diarrhea samples and 12 ascites samples). The coincidence rate between FCoV-RPA and RT-qPCR for detection in clinical samples was 95.2%. In summary, FCoV-RPA analysis provides an efficient, rapid, and sensitive detection method for FCoV.

Keywords: Detection; Feline coronavirus; Recombinase polymerase amplification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cat Diseases / diagnosis
  • Cat Diseases / virology
  • Cats
  • Coronavirus Infections / diagnosis*
  • Coronavirus, Feline / genetics*
  • Coronavirus, Feline / isolation & purification
  • Feline Infectious Peritonitis / diagnosis*
  • Molecular Diagnostic Techniques / methods
  • Molecular Diagnostic Techniques / veterinary*
  • Nucleic Acid Amplification Techniques / methods*
  • Nucleic Acid Amplification Techniques / veterinary
  • RNA, Viral / genetics*
  • Real-Time Polymerase Chain Reaction / methods
  • Sensitivity and Specificity

Substances

  • RNA, Viral

Supplementary concepts

  • LAMP assay