{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T19:45:14Z","timestamp":1789847114142,"version":"4.0.1"},"reference-count":71,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T00:00:00Z","timestamp":1764892800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005621","name":"Kasetsart University Research and Development Institute, KURDI","doi-asserted-by":"crossref","award":["FF(KU-SRIU)7.68"],"award-info":[{"award-number":["FF(KU-SRIU)7.68"]}],"id":[{"id":"10.13039\/501100005621","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100022435","name":"Kasetsart University Institute for Advanced Studies","doi-asserted-by":"crossref","award":["RUP69_Act2_No.17 (PD)"],"award-info":[{"award-number":["RUP69_Act2_No.17 (PD)"]}],"id":[{"id":"10.13039\/501100022435","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100022271","name":"Faculty of Veterinary Medicine, Kasetsart University","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100022271","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Feline leukemia virus (FeLV) is a retrovirus that globally affects both domestic and wild cats, leading to the development of leukemia, lymphoma, and immunosuppression. However, it is important to note that the daily antigen test may yield false negative results. In this study, we successfully developed the first colorimetric loop-mediated isothermal amplification (LAMP) associated with neutral red (NR-LAMP) for the detection of FeLV. The NR-LAMP assay exhibited high sensitivity and efficiency compared to the routine polymerase chain reaction (PCR) reference method. To ensure specificity, a novel LAMP primer set was custom-designed based on the pol gene of multiple FeLV strains, which resulted in no cross-amplification with other feline viruses. Under the optimized isothermal amplification conditions at 61 \u00b0C for 40 min, the NR-LAMP assay achieved a detection limit of 100 copies\/\u00b5L. Using a blind clinical test involving 98 samples, the NR-LAMP assay demonstrated perfect agreement with the reference PCR method, providing a sensitivity of 97.3% and a specificity of 100%. This proposed NR-LAMP assay surpasses other related approaches in terms of sensitivity, efficiency, and cost-effectiveness. Consequently, the colorimetric NR-LAMP reaction serves as a robust and convenient diagnostic tool for the inspection of FeLV, offering an alternative molecular method for future clinical applications and commercial utilization.<\/jats:p>","DOI":"10.3390\/ijms262411793","type":"journal-article","created":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T16:37:02Z","timestamp":1764952622000},"page":"11793","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Innovative Colorimetric Neutral Red-Based Loop-Mediated Isothermal Amplification (NR-LAMP) Assay: Transforming Rapid and Affordable Feline Leukemia Virus Detection"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-8529-4137","authenticated-orcid":false,"given":"Witsanu","family":"Rapichai","sequence":"first","affiliation":[{"name":"Center for Advanced Studies for Agriculture and Food, Kasetsart University Institute for Advanced Studies, Kasetsart University, Bangkok 10900, Thailand"},{"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":[{"vocabulary":"crossref","role":"author"}]},{"given":"Piyamat","family":"Khamsingnok","sequence":"additional","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anyalak","family":"Wachirachaikarn","sequence":"additional","affiliation":[{"name":"Department of Sciences and Bioinnovation, Faculty of Liberal Arts and Science, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, 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Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"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":[{"vocabulary":"crossref","role":"author"}]},{"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, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Supansa","family":"Tuanthap","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine, Rajamangala University of Technology Tawan-ok, Bangpra, Chonburi 20110, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amonpun","family":"Rattanasrisomporn","sequence":"additional","affiliation":[{"name":"Interdisciplinary of Genetic Engineering and Bioinformatics, Graduate School, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Selapoom","family":"Pairor","sequence":"additional","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2421-7859","authenticated-orcid":false,"given":"Kiattawee","family":"Choowongkomon","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, 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