{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:30:10Z","timestamp":1760059810211,"version":"build-2065373602"},"reference-count":59,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T00:00:00Z","timestamp":1752710400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Feline coronavirus (FCoV) is a major pathogen causing feline infectious peritonitis (FIP), a lethal disease in cats, necessitating accurate diagnostic methods. This study developed and compared novel primers targeting the FCoV membrane (M) gene for enhanced detection. Specific primers were designed for the M gene and their performance evaluated using reverse transcription-PCR (RT-PCR), nested RT-PCR, and reverse transcription-quantitative PCR (RT-qPCR) on 80 clinical effusion samples from cats suspected of FIP. Specificity of assays was tested against other feline viruses, with sensitivity being assessed via serial dilutions of FCoV RNA. RT-qPCR had the highest sensitivity, detecting 9.14 \u00d7 101 copies\/\u00b5L, identifying 93.75% of positive samples, followed by nested RT-PCR (87.50%, 9.14 \u00d7 104 copies\/\u00b5L) and RT-PCR (61.25%, 9.14 \u00d7 106 copies\/\u00b5L). All assays had 100% specificity, with no cross-reactivity to other viruses. The nested RT-PCR and RT-qPCR outperformed RT-PCR significantly, with comparable diagnostic accuracy. The novel primers targeting the FCoV M gene, coupled with RT-qPCR, delivered unparalleled sensitivity and robust reliability for detecting FCoV in clinical settings. Nested RT-PCR was equally precise and amplified diagnostic confidence with its high performance. These cutting-edge assays should revolutionize FCoV detection, offering trusted tools that seamlessly integrate into veterinary practice, empowering clinicians to manage feline infectious peritonitis with unprecedented accuracy and speed.<\/jats:p>","DOI":"10.3390\/ijms26146861","type":"journal-article","created":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T11:25:17Z","timestamp":1752751517000},"page":"6861","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Detection of Feline Coronavirus Membrane Gene Based on Conventional Revere Transcription-Polymerase Chain Reaction, Nested Reverse Transcription-Polymerase Chain Reaction, and Reverse Transcription-Quantitative Polymerase Chain Reaction: A Comparative Study"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-7030-8953","authenticated-orcid":false,"given":"Chiraphat","family":"Kopduang","sequence":"first","affiliation":[{"name":"Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8529-4137","authenticated-orcid":false,"given":"Witsanu","family":"Rapichai","sequence":"additional","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-5066-0041","authenticated-orcid":false,"given":"Chalandhorn","family":"Leangcharoenpong","sequence":"additional","affiliation":[{"name":"Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Piyamat","family":"Khamsingnok","sequence":"additional","affiliation":[{"name":"Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thanapol","family":"Puangmalee","sequence":"additional","affiliation":[{"name":"Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1078-186X","authenticated-orcid":false,"given":"Siriluk","family":"Ratanabunyong","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amonpun","family":"Rattanasrisomporn","sequence":"additional","affiliation":[{"name":"Interdisciplinary of Genetic Engineering and Bioinformatics, Graduate School, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thanawat","family":"Khaoiam","sequence":"additional","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Kasetsart University Veterinary Teaching Hospital, Faculty of Veterinary Medicine, Kamphaeng Saen Campus, Kasetsart University, Nakhon Pathom 73140, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6340-9204","authenticated-orcid":false,"given":"Hieu Van","family":"Dong","sequence":"additional","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Trau Quy Town, Gia Lam District, Hanoi 131000, Vietnam"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2421-7859","authenticated-orcid":false,"given":"Kiattawee","family":"Choowongkomol","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2426-0961","authenticated-orcid":false,"given":"Jatuporn","family":"Rattanasrisomporn","sequence":"additional","affiliation":[{"name":"Graduate Program in Animal Health and Biomedical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, 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