{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T13:34:55Z","timestamp":1791725695665,"version":"4.3.4"},"reference-count":74,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,5,22]],"date-time":"2023-05-22T00:00:00Z","timestamp":1684713600000},"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. Bioeng. Biotechnol."],"abstract":"<jats:p><jats:bold>Introduction:<\/jats:bold> Bacterial wilt (BW) caused by the aerobic, Gram-negative pathogenic species <jats:italic>Ralstonia solanacearum<\/jats:italic> (RS) is a major disease impacting commercial agriculture worldwide. Asian phylotype I of RS is the cause of tomato bacterial wilt, which has caused severe economic losses in southern China for many years. An urgent priority in control of bacterial wilt is development of rapid, sensitive, effective methods for detection of RS.<\/jats:p><jats:p><jats:bold>Methods:<\/jats:bold> We describe here a novel RS detection assay based on combination of loop-mediated isothermal amplification (LAMP) and CRISPR\/Cas12a. crRNA1, with high trans-cleavage activity targeting <jats:italic>hrpB<\/jats:italic> gene, was selected out of four candidate crRNAs. Two visual detection techniques, involving naked-eye observation of fluorescence and lateral flow strips, were tested and displayed high sensitivity and strong specificity.<\/jats:p><jats:p><jats:bold>Results and Discussion:<\/jats:bold> The LAMP\/Cas12a assay accurately detected RS phylotype \u2160 in 14 test strains, and showed low detection limit (2.0 \u00d7 10<jats:sup>0<\/jats:sup> copies). RS in tomato stem tissue and soil samples from two field sites with suspected BW infection was identified accurately, suggesting potential application of LAMP\/Cas12a assay as point-of-care test (POCT). The overall detection process took less than 2 h and did not require professional lab equipment. Our findings, taken together, indicate that LAMP\/Cas12a assay can be developed as an effective, inexpensive technique for field detection and monitoring of RS.<\/jats:p>","DOI":"10.3389\/fbioe.2023.1188176","type":"journal-article","created":{"date-parts":[[2023,5,22]],"date-time":"2023-05-22T04:46:55Z","timestamp":1684730815000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":16,"title":["Loop-mediated isothermal amplification (LAMP)\/Cas12a assay for detection of Ralstonia solanacearum in tomato"],"prefix":"10.3389","volume":"11","author":[{"given":"Zhiyu","family":"Fan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxia","family":"Mei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiawei","family":"Xing","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tian","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Di","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingjun","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Derui","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zufeng","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunxiang","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,5,22]]},"reference":[{"key":"B1","first-page":"126","article-title":"Culture media for Ralstonia solanacearum isolation, identification and maintenance","volume":"30","author":"Aley","year":"1995","journal-title":"Fitopatologia"},{"key":"B2","doi-asserted-by":"publisher","first-page":"5655","DOI":"10.1128\/jb.179.18.5655-5662.1997","article-title":"The type III (hrp) secretion pathway of plant pathogenic bacteria: Trafficking harpins, avr proteins, and death","volume":"179","author":"Alfano","year":"1997","journal-title":"J. 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