{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T07:17:59Z","timestamp":1788419879941,"version":"build-2803163510"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Newcastle Disease Virus (NDV) is not only a significant and persistent threat to the healthy development of poultry industry, but also an important pathogen endangering food safety and harboring potential zoonotic risks.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>In this study, a rapid and highly sensitive diagnostic assay was established for genotype VII NDV by integrating reverse transcription recombinase polymerase amplification (RT-RPA) with the clustered regularly interspaced short palindromic repeats (CRISPR)\/Cas13a system, in combination with a lateral flow dipstick (LFD) for visual signal readout. Specific RT-RPA primers were designed based on the fusion (F) gene of genotype VII NDV, and CRISPR RNAs (crRNAs) were constructed to establish a simplified CRISPR\/Cas13a-based diagnostic workflow targeting the fusion (F) gene of genotype VII NDV.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The optimized detection platform integrates RT-RPA amplification at 39\u202f\u00b0C for 20\u202fmin and CRISPR\/Cas13a-mediated cleavage at 37\u202f\u00b0C for 20\u202fmin. The final results can be quantified by fluorescence signals or visualized via LFD band within 5\u202fmin, with the total detection procedure completed in less than 45\u202fmin. Performance evaluation demonstrated that this assay achieved a limit of detection (LOD) as low as 6 copies per microliter of input and high specificity, with no cross-reactivity against other common avian pathogens. Clinical validation of 154 samples showed 100% concordance between the RT-RPA-CRISPR\/Cas13a-(LFD) assay and RT-PCR sequencing for genotype VII NDV detection.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>This method requires no sophisticated instruments and enables simple operation. It provides a field-deployable tool for rapid on-site detection of genotype VII NDV, which is suitable for preliminary screening in primary laboratories, and holds promising application potential for the early surveillance and food safety monitoring of virulent NDV strains.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fvets.2026.1870607","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T07:10:37Z","timestamp":1788419437000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["On-site and visual detection of genotype VII Newcastle disease virus based on RT-RPA and CRISPR\/Cas13a"],"prefix":"10.3389","volume":"13","author":[{"given":"Yaping","family":"Huang","sequence":"first","affiliation":[{"name":"Minjiang Teachers College","place":["Fuzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Animal Husbandry and Veterinary Medicine\/Fujian Key Laboratory for Control and Prevention of Avian Diseases, Fujian Academy of Agricultural Sciences","place":["Fuzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhui","family":"Song","sequence":"additional","affiliation":[{"name":"College of Bee Science and Biomedicine, Fujian Agriculture and Forestry University","place":["Fuzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qizhang","family":"Liang","sequence":"additional","affiliation":[{"name":"Institute of Animal Husbandry and Veterinary Medicine\/Fujian Key Laboratory for Control and Prevention of Avian Diseases, Fujian Academy of Agricultural Sciences","place":["Fuzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiuling","family":"Fu","sequence":"additional","affiliation":[{"name":"Institute of Animal Husbandry and Veterinary Medicine\/Fujian Key Laboratory for Control and Prevention of Avian Diseases, Fujian Academy of Agricultural Sciences","place":["Fuzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ningning","family":"Cheng","sequence":"additional","affiliation":[{"name":"College of Animal Sciences, Fujian Agriculture and Forestry University","place":["Fuzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Xie","sequence":"additional","affiliation":[{"name":"Minjiang Teachers College","place":["Fuzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guanghua","family":"Fu","sequence":"additional","affiliation":[{"name":"Institute of Animal Husbandry and Veterinary Medicine\/Fujian Key Laboratory for Control and Prevention of 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