Clinical evaluation of a direct RT-qPCR method for feline coronavirus detection in effusions using the PicoGene PCR1100 system

Abstract

Feline coronavirus (FCoV) infects both domestic and wild felids and has the potential to cause feline infectious peritonitis (FIP), a progressive and often fatal systemic disease. Although rapid diagnosis and treatment are crucial in cases of FIP, conventional reverse-transcription quantitative real-time PCR (RT-qPCR) requires RNA extraction and specialized equipment, limiting its use for timely testing in general veterinary practice. We evaluated the performance of a direct RT-qPCR method using the PicoGene PCR1100 system (GoFoton, Ibaraki, Japan), which omits the RNA extraction step and delivers results within ~40 min. Compared with FCoV culture supernatants and extracted RNA, we estimated the limit of detection of this direct RT-qPCR method to be 150 copies/reaction—a detection sensitivity equivalent to that of conventional RT-qPCR targeting the FCoV 3′-UTR. We observed no cross-reactivity with other feline viruses or SARS-CoV-2. We subsequently analyzed 28 pleural and abdominal effusions collected from cats suspected of having FIP to compare the direct RT-qPCR method with the conventional approach. The sensitivity of the direct RT-qPCR method was 95.5% (95% CI: [78.2, 99.2]) and the specificity was 100% (95% CI: [61.0, 100.0]), which supports the use of the PCR1100 system as a rapid and user-friendly point-of-care tool for the detection of FCoV RNA in effusion samples.