{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T15:36:11Z","timestamp":1791300971001,"version":"4.1.0"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,4,28]],"date-time":"2023-04-28T00:00:00Z","timestamp":1682640000000},"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. Public Health"],"abstract":"<jats:p>Melioidosis is a bacterial infection caused by <jats:italic>Burkholderia pseudomallei<\/jats:italic> (<jats:italic>B. pseudomallei<\/jats:italic>), posing a significant threat to public health. Rapid and accurate detection of <jats:italic>B. pseudomallei<\/jats:italic> is crucial for preventing and controlling melioidosis. However, identifying <jats:italic>B. pseudomallei<\/jats:italic> is challenging due to its high similarity to other species in the same genus. To address this issue, this study proposed a dual-target method that can specifically identify <jats:italic>B. pseudomallei<\/jats:italic> in less than 40\u2009min. We analyzed 1722 <jats:italic>B. pseudomallei<\/jats:italic> genomes to construct large-scale pan-genomes and selected specific sequence tags in their core genomes that effectively distinguish <jats:italic>B. pseudomallei<\/jats:italic> from its closely related species. Specifically, we selected two specific tags, LC1 and LC2, which we combined with the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated proteins (Cas12a) system and recombinase polymerase amplification (RPA) pre-amplification. Our analysis showed that the dual-target RPA-CRISPR\/Cas12a assay has a sensitivity of approximately 0.2 copies\/reaction and 10\u2009fg genomic DNA for LC1, and 2 copies\/reaction and 20\u2009fg genomic DNA for LC2. Additionally, our method can accurately and rapidly detect <jats:italic>B. pseudomallei<\/jats:italic> in human blood and moist soil samples using the specific sequence tags mentioned above. In conclusion, the dual-target RPA-CRISPR\/Cas12a method is a valuable tool for the rapid and accurate identification of <jats:italic>B. pseudomallei<\/jats:italic> in clinical and environmental samples, aiding in the prevention and control of melioidosis.<\/jats:p>","DOI":"10.3389\/fpubh.2023.1153352","type":"journal-article","created":{"date-parts":[[2023,5,5]],"date-time":"2023-05-05T05:06:08Z","timestamp":1683263168000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":12,"title":["Detection of Burkholderia pseudomallei with CRISPR-Cas12a based on specific sequence 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