{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T13:58:57Z","timestamp":1790344737870,"version":"4.1.0"},"reference-count":32,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:00:00Z","timestamp":1790294400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:00:00Z","timestamp":1790294400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"Open Research Project of Zhejiang Key Laboratory of Public Health Detection and Pathogenesis Research","award":["2024KF22"],"award-info":[{"award-number":["2024KF22"]}]},{"name":"Zhejiang Science and Technology Plan for Disease Prevention and Control","award":["2025JK141, 2025JKY0458"],"award-info":[{"award-number":["2025JK141, 2025JKY0458"]}]},{"name":"Mindray Joint Fund of Zhejiang Provincial Natural Science Foundation of China","award":["LMRY26H200007"],"award-info":[{"award-number":["LMRY26H200007"]}],"id":[{"id":"https:\/\/ror.org\/04aqat463","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Burkholderia gladioli<\/jats:italic>\n                      pathovar\n                      <jats:italic toggle=\"yes\">cocovenenans<\/jats:italic>\n                      produces the potent mitochondrial toxin bongkrekic acid (BA), causing severe foodborne outbreaks. Conventional detection methods are constrained by prolonged turnaround times and the need for sophisticated instruments, limiting their utility for on-site surveillance. In this study, two independent one-step RPA-CRISPR\/Cas13a detection systems were established: one targeting a species-level marker for broad screening of\n                      <jats:italic toggle=\"yes\">B. gladioli<\/jats:italic>\n                      and the other targeting the\n                      <jats:italic toggle=\"yes\">bonA<\/jats:italic>\n                      gene cluster for specific identification of the toxigenic pathovar. The complete one-step reaction was performed at 37\u00b0C for 30 min, and results were visually interpreted using a portable blue-light transilluminator. The visual detection limit reached 1 \u00d7 10\u2070 copy\/\u03bcL for both assays, with no cross-reactivity observed against a panel of six non-target strains. Component validation experiments confirmed that all listed constituents were indispensable for fluorescence signal generation. The practical applicability of the platform was evaluated using 100 naturally contaminated black fungus and mushroom substrate samples. The species-level assay returned positive rates of 78% (39\/50) for black fungus and 42% (21\/50) for substrate samples, while the\n                      <jats:italic toggle=\"yes\">bonA<\/jats:italic>\n                      -targeted assay yielded positive rates of 38% (19\/50) and 8% (4\/50), respectively. The visual readout results were in 100% concordance with conventional quantitative real-time PCR. This field-deployable, equipment-free visual detection platform holds considerable promise for on-site surveillance and early warning of BA-producing\n                      <jats:italic toggle=\"yes\">B. gladioli<\/jats:italic>\n                      in food and environmental matrices.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Bongkrekic acid (BA) poisoning caused by\n                        <jats:italic toggle=\"yes\">Burkholderia gladioli<\/jats:italic>\n                        pv.\n                        <jats:italic toggle=\"yes\">cocovenenans<\/jats:italic>\n                        is a lethal food safety threat, particularly in China and Southeast Asia. Current detection methods depend heavily on laboratory-based instruments, limiting their utility for on-site surveillance. This study developed two independent one-step RPA-CRISPR\/Cas13a detection systems: one for broad screening of\n                        <jats:italic toggle=\"yes\">B. gladioli<\/jats:italic>\n                        at the species level and the other for specific identification of the toxigenic pathovar. The one-step reaction was completed at 37\u00b0C within 30 min, and results were visually interpreted using only a portable blue-light transilluminator. The visual detection limit reached 1 \u00d7 10\u2070 copy\/\u03bcL, and the method exhibited 100% concordance with quantitative real-time PCR when applied to 100 naturally contaminated samples. This field-deployable, equipment-free platform provides a practical tool for on-site surveillance and early warning of BA-producing\n                        <jats:italic toggle=\"yes\">B. gladioli<\/jats:italic>\n                        in food and environmental matrices.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.02143-26","type":"journal-article","created":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T13:00:18Z","timestamp":1790341218000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Field-deployable visual detection of\n                    <i>Burkholderia gladioli<\/i>\n                    and its toxigenic pathovar using a one-step RPA-CRISPR\/Cas13a assay"],"prefix":"10.1128","author":[{"given":"Jinkai","family":"Li","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04epb4p87","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Medical Technology, Zhejiang Chinese Medical University","place":["Hangzhou, China"]},{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Gong","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaomeng","family":"Huang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fuming","family":"Liu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianhua","family":"Mei","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siyu","family":"Fu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liyang","family":"Qiu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-3884-9650","authenticated-orcid":false,"given":"Deyong","family":"Zhang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2098-2817","authenticated-orcid":true,"given":"Dongqing","family":"Cheng","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04epb4p87","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Medical Technology, Zhejiang Chinese Medical University","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8091-4288","authenticated-orcid":true,"given":"Xiuying","family":"Chen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02x2tv943","id-type":"ROR","asserted-by":"publisher"}],"name":"Lishui Center for Disease Control and Prevention","place":["Lishui, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2026,9,25]]},"reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijid.2022.05.012"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s13181-016-0577-1"},{"key":"e_1_3_3_4_2","unstructured":"WHO. 2015. WHO estimates of the global burden of foodborne diseases: foodborne disease burden epidemiology reference group 2007\u20132015. Geneva Switzerland World Health Organization. https:\/\/www.who.int\/publications\/i\/item\/9789241565165."},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cofs.2021.01.006"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cix1005"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X.JFP-19-121"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.46234\/ccdcw2020.264"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.fm.2023.104249"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.3390\/foods12213926"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-03458-6"},{"key":"e_1_3_3_12_2","unstructured":"National Health Commission of the People\u2019s Republic of China. 2020. National food safety standard\u2014food microbiological examination\u2014Burkholderia gladioli (Pseudomonas cocovenenans subsp. farinfermentans). GB 4789.29-2020. National Health Commission of the People\u2019s Republic of China. Beijing China . https:\/\/www.chinesestandard.net\/PDF.aspx\/GB4789.29-2020#ViewPDF."},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.02677"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.abi9335"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41551-021-00760-7"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.snb.2019.127440"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2017.05.008"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.10.044"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41421-018-0028-z"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature24049"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf5573"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaq0179"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/acssensors.8b01206"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2015.245829"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar6245"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-17994-9"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aam9321"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkae973"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.13590\/j.cjfh.2022.06.035"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2012.07.022"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1080\/87559129.2023.2199829"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1080\/14737159.2022.2109964"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.06.050"}],"container-title":["Microbiology Spectrum"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.02143-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.02143-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T13:00:20Z","timestamp":1790341220000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.02143-26"}},"subtitle":[],"editor":[{"given":"Dan","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,25]]},"references-count":32,"alternative-id":["10.1128\/spectrum.02143-26"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.02143-26","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,25]]},"assertion":[{"value":"2026-06-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-19","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-25","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e02143-26"}}