Development of a recombinase polymerase amplification fluorescence assay to detect feline coronavirus. (December 2020)

• Record Type: Journal Article
• Authors: Hu, Xiaoliang; Xiao, Li; Cong, Xiao; Zhu, Yujun; Huang, Bihong; Cong, Feng
• Is Part Of: Molecular and cellular probes. Volume 54(2020)
• Publication Date: 2020-12
• Subjects: Recombinase polymerase amplification -- Feline coronavirus -- Detection; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2020.101669
• Languages: English
• ISSNs: 0890-8508

Abstract

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. Highlights: FCoV RPA has the shortest reaction time within 15 min among all the PCR-based methods. FCoV RPA detection method has no cross-reactivity with other pathogens. The sensitivity of FCoV RPA was consistent with real-time PCR, as low as 204 copies/μL of DNA molecules per reaction.