{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T05:55:01Z","timestamp":1781848501070,"version":"3.54.5"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T00:00:00Z","timestamp":1781827200000},"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. Cell. Infect. Microbiol."],"abstract":"\n Background<\/jats:title>\n \n Foodborne illnesses caused by enterotoxigenic\n Bacillus cereus<\/jats:italic>\n represent a significant public health concern globally. Rapid and simple detection methods are needed, particularly in resource-limited settings where conventional laboratory infrastructure is lacking. This study aimed to develop and validate a loop-mediated isothermal amplification (LAMP) assay targeting three major enterotoxin genes (\n nheA<\/jats:italic>\n ,\n cytK<\/jats:italic>\n , and\n entFM<\/jats:italic>\n ) for the rapid detection of enterotoxigenic\n B<\/jats:italic>\n .\n cereus<\/jats:italic>\n in food and feed samples from Bangladesh.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n LAMP primers for the\n nheA<\/jats:italic>\n gene were newly designed, while\n cytK<\/jats:italic>\n and\n entFM<\/jats:italic>\n primers were adapted from previous studies. Reaction conditions were optimized using\n B<\/jats:italic>\n .\n cereus<\/jats:italic>\n ATCC 14579. Analytical sensitivity was determined using serial dilutions of pure culture, spiked milk, and genomic DNA. Specificity was assessed against 20 bacterial strains, including\n Bacillus<\/jats:italic>\n species and non-\n Bacillus<\/jats:italic>\n foodborne pathogens. The assay was validated on 30 field samples (feed, milk, and eggs) and compared with conventional PCR.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Optimal LAMP amplification was achieved at 64\u00b0C for 40\u00a0min. The limit of detection was 100 fg\/\u00b5L for genomic DNA, 9 \u00d7 10\n 2<\/jats:sup>\n CFU\/mL for pure culture, and 9 \u00d7 10\n 3<\/jats:sup>\n CFU\/mL for spiked milk\u2014approximately 10,000-fold more sensitive than conventional PCR. The assay showed 100% specificity for enterotoxigenic\n Bacillus<\/jats:italic>\n species, with no cross-amplification of non-target bacteria. In field samples, LAMP detected enterotoxin genes in 73.3% (22\/30) for\n nheA<\/jats:italic>\n , 53.3% (16\/30) for\n cytK<\/jats:italic>\n , and 80.0% (24\/30) for\n entFM<\/jats:italic>\n , compared to 63.3% (19\/30), 40.0% (12\/30), and 70.0% (21\/30) by PCR, respectively. Overall diagnostic sensitivity and specificity against PCR were 96.1% and 66.7%, respectively, with substantial agreement (\u03ba\u00a0= 0.62).\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n Although LAMP assays for individual\n B<\/jats:italic>\n .\n cereus<\/jats:italic>\n toxin genes have been described previously, this study represents the first application of this specific\n nheA<\/jats:italic>\n -\n cytK<\/jats:italic>\n -\n entFM<\/jats:italic>\n LAMP panel in the Bangladeshi food and feed context, providing a regionally relevant, field-deployable method for food safety monitoring.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1815090","type":"journal-article","created":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T05:48:35Z","timestamp":1781848115000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Rapid detection of enterotoxigenic Bacillus cereus by loop-mediated isothermal amplification assay from food and feed samples"],"prefix":"10.3389","volume":"16","author":[{"given":"Md Atiqul","family":"Haque","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University","place":["Dinajpur, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Arifur","family":"Rahman","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Noakhali Science and Technology University","place":["Noakhali, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Israt","family":"Islam","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Noakhali Science and Technology University","place":["Noakhali, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uditi Paul","family":"Bristi","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Noakhali Science and Technology University","place":["Noakhali, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farzana","family":"Islam","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Noakhali Science and Technology University","place":["Noakhali, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nazmul","family":"Alam","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Noakhali Science and Technology University","place":["Noakhali, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanjina","family":"Sultana","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, 250 Bedded General Hospital","place":["Noakhali, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jubayer","family":"Mumin","sequence":"additional","affiliation":[{"name":"Department of Global Public Health, Karolinska Institute","place":["Stockholm, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Firoz","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, Noakhali Science and Technology University","place":["Noakhali, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng","family":"He","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Veterinary Public Health, College of Veterinary Medicine, China Agricultural University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Aminul","family":"Islam","sequence":"additional","affiliation":[{"name":"Advanced Molecular Lab, Department of Microbiology, President Abdul Hamid Medical College","place":["Kishoreganj, Bangladesh"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,6,19]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1630","DOI":"10.38124\/IJISRT20AUG795","article-title":"Epidemiological investigation, prevalence & antibiogram study of potential zoonotic bacterial pathogens of household pets at Dinajpur district of Bangladesh","volume":"5","author":"Akter","year":"2020","journal-title":"Int. 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