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Currently, there are no effective vaccines or treatments for ALV. Therefore, the early diagnosis of infected flocks and farm sanitation are crucial for controlling outbreaks of this disease. To address the limitations of traditional diagnostic methods, which require sophisticated equipment and skilled personnel, a dual-tube detection method for ALV-J based on reverse transcription isothermal amplification (RAA) and the CRISPR-Cas13a system has been developed. This method offers the advantages of high sensitivity, specificity, and rapidity; it is capable of detecting virus concentrations as low as 5.4 \u00d7 100 copies\/\u03bcL without cross-reactivity with other avian viruses, with a total testing time not exceeding 85 min. The system was applied to 429 clinical samples, resulting in a positivity rate of 15.2% for CRISPR-Cas13a, which was higher than the 14.7% detected by PCR and 14.2% by ELISA, indicating superior detection capability and consistency. Furthermore, the dual-tube RAA-CRISPR detection system provides visually interpretable results, making it suitable for on-site diagnosis in remote farms lacking laboratory facilities. In conclusion, the proposed ALV-J detection method, characterized by its high sensitivity, specificity, and convenience, is expected to be a vital technology for purification efforts against ALV-J.<\/jats:p>","DOI":"10.3390\/ijms251910780","type":"journal-article","created":{"date-parts":[[2024,10,7]],"date-time":"2024-10-07T10:58:32Z","timestamp":1728298712000},"page":"10780","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Detection of Avian Leukosis Virus Subgroup J (ALV-J) Using RAA and CRISPR-Cas13a Combined with Fluorescence and Lateral Flow Assay"],"prefix":"10.3390","volume":"25","author":[{"given":"Shutao","family":"Chen","sequence":"first","affiliation":[{"name":"Chongqing Key Laboratory of Herbivore Science, College of Animal Science and Technology, Southwest University, Chongqing 400700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhang","family":"Li","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of Herbivore Science, College of Animal Science and Technology, Southwest University, Chongqing 400700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruyu","family":"Liao","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of Herbivore Science, College of Animal Science and Technology, Southwest University, Chongqing 400700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng","family":"Liu","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of Herbivore Science, College of Animal Science and Technology, Southwest University, Chongqing 400700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyi","family":"Zhou","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of Herbivore Science, College of Animal Science and Technology, Southwest University, Chongqing 400700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiwei","family":"Wang","sequence":"additional","affiliation":[{"name":"Chongqing Academy of Animal Science, Chongqing 402460, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qigui","family":"Wang","sequence":"additional","affiliation":[{"name":"Chongqing Academy of Animal Science, Chongqing 402460, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-1203-2059","authenticated-orcid":false,"given":"Xi","family":"Lan","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of Herbivore Science, College of Animal Science and Technology, Southwest University, Chongqing 400700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,7]]},"reference":[{"key":"ref_1","first-page":"1167","article-title":"Myeloid leukaemogenicity and transmission of the HPRS-103 strain of avian leukosis virus","volume":"6","author":"Payne","year":"1992","journal-title":"Leukemia"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1080\/03079457.2011.646237","article-title":"The long view: 40 years of avian leukosis research","volume":"41","author":"Payne","year":"2012","journal-title":"Avian Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1080\/03079450400025356","article-title":"Single and concurrent avian leukosis virus infections with avian leukosis virus-J and avian leukosis virus-A in Australian meat-type chickens","volume":"34","author":"Fenton","year":"2005","journal-title":"Avian Pathol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"604","DOI":"10.2307\/1592664","article-title":"Body weight suppression in broilers naturally infected with avian leukosis virus subgroup J","volume":"43","author":"Stedman","year":"1999","journal-title":"Avian Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1099\/0022-1317-72-4-801","article-title":"A novel subgroup of exogenous avian leukosis virus in chickens","volume":"72","author":"Payne","year":"1991","journal-title":"J. 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