{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T05:23:16Z","timestamp":1787289796599,"version":"build-2736575974"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T00:00:00Z","timestamp":1787270400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009110","name":"Natural Science Foundation of Xinjiang Uygur Autonomous Region","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100009110","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>\n                    Bovine babesiosis, caused by\n                    <jats:italic>Babesia bovis<\/jats:italic>\n                    , is a tick-borne hemoparasitic disease causing substantial economic losses to cattle industries worldwide. Current diagnostic methods, particularly microscopy, lack sensitivity for subclinical infections, while molecular assays require sophisticated equipment unsuitable for field use. To address this gap, we developed a novel diagnostic assay integrating enzymatic recombinase amplification (ERA) with CRISPR-Cas12a-mediated trans-cleavage, targeting the\n                    <jats:italic>B. bovis<\/jats:italic>\n                    spherical body protein 2 (SBP2) gene. The assay enables visual readout via UV\/blue light fluorescence and lateral flow strips. Following systematic optimization of primers, crRNA, reaction temperature, and reporter concentration, the ERA-CRISPR\/Cas12a system achieved complete amplification within 20 min at 37 \u00b0C and demonstrated a detection limit of 10 copies\/\u03bcL with high specificity for\n                    <jats:italic>B. bovis<\/jats:italic>\n                    . comparable diagnostic performance with conventional PCR within the tested sample panel. The fluorescence readout achieved a detection limit of 10\n                    <jats:sup>1<\/jats:sup>\n                    copies\/\u03bcL, while the lateral-flow strip format exhibited a detection limit of 104 copies\/\u03bcL. The entire procedure, from DNA extraction (approximately 30 min) to final result readout (ERA: 20 min; CRISPR-Cas12a: 20 min), can be completed within approximately 70 min. This detection method supports two types of visual detection: portable blue light and test strips, and is suitable for on-site environments with limited resources. This work provides a practical diagnostic tool for rapid, accurate detection of bovine babesiosis, with potential applicability to other veterinary pathogens.\n                  <\/jats:p>","DOI":"10.3389\/fvets.2026.1917943","type":"journal-article","created":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T04:22:55Z","timestamp":1787286175000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of an ERA-CRISPR\/Cas12a-based visual diagnostic assay for bovine babesiosis"],"prefix":"10.3389","volume":"13","author":[{"given":"Yongchang","family":"Li","sequence":"first","affiliation":[{"name":"Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University","place":["Urumqi, Xinjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiujuan","family":"Feng","sequence":"additional","affiliation":[{"name":"Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University","place":["Urumqi, Xinjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianyue","family":"Fu","sequence":"additional","affiliation":[{"name":"Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University","place":["Urumqi, Xinjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fakiha","family":"Kalim","sequence":"additional","affiliation":[{"name":"Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University","place":["Urumqi, Xinjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiyun","family":"Li","sequence":"additional","affiliation":[{"name":"Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University","place":["Urumqi, Xinjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iqra","family":"Zafar","sequence":"additional","affiliation":[{"name":"Laboratory of Sustainable Animal Environment, Graduate School of Agriculture Science, Tohoku University","place":["Sendai, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed Abdo","family":"Rizk","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University","place":["Mansoura, Egypt"]},{"name":"Department of Clinical Science, College of Veterinary Medicine, King Faisal University","place":["Al-Hofuf, Al-Ahsa, Saudi Arabia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeyun","family":"Cui","sequence":"additional","affiliation":[{"name":"Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University","place":["Urumqi, Xinjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bayin","family":"Chahan","sequence":"additional","affiliation":[{"name":"Parasitology Laboratory, Veterinary College, Xinjiang Agricultural University","place":["Urumqi, Xinjiang, 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