{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T22:32:19Z","timestamp":1778711539307,"version":"3.51.4"},"reference-count":35,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2001,12,1]],"date-time":"2001-12-01T00:00:00Z","timestamp":1007164800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2001,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Botulism is diagnosed by detecting botulinum neurotoxin and<jats:italic>Clostridium botulinum<\/jats:italic>cells in the patient and in suspected food samples. In this study, a multiplex PCR assay for the detection of<jats:italic>Clostridium botulinum<\/jats:italic>types A, B, E, and F in food and fecal material was developed. The method employs four new primer pairs with equal melting temperatures, each being specific to botulinum neurotoxin gene type A, B, E, or F, and enables a simultaneous detection of the four serotypes. A total of 43<jats:italic>C. botulinum<\/jats:italic>strains and 18 strains of other bacterial species were tested. DNA amplification fragments of 782 bp for<jats:italic>C. botulinum<\/jats:italic>type A alone, 205 bp for type B alone, 389 bp for type E alone, and 543 bp for type F alone were obtained. Other bacterial species, including<jats:italic>C. sporogenes<\/jats:italic>and the nontoxigenic nonproteolytic<jats:italic>C. botulinum-<\/jats:italic>like organisms, did not yield a PCR product. Sensitivity of the PCR for types A, E, and F was 10<jats:sup>2<\/jats:sup>cells and for type B was 10 cells per reaction mixture. With a two-step enrichment, the detection limit in food and fecal samples varied from 10<jats:sup>\u22122<\/jats:sup>spore\/g for types A, B, and F to 10<jats:sup>\u22121<\/jats:sup>spore\/g of sample material for type E. Of 72 natural food samples investigated, two were shown to contain<jats:italic>C. botulinum<\/jats:italic>type A, two contained type B, and one contained type E. The assay is sensitive and specific and provides a marked improvement in the PCR diagnostics of<jats:italic>C. botulinum<\/jats:italic>.<\/jats:p>","DOI":"10.1128\/aem.67.12.5694-5699.2001","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:03:59Z","timestamp":1027764239000},"page":"5694-5699","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":140,"title":["Multiplex PCR Assay for Detection and Identification of<i>Clostridium botulinum<\/i>Types A, B, E, and F in Food and Fecal Material"],"prefix":"10.1128","volume":"67","author":[{"given":"Miia","family":"Lindstro\u0308m","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, FIN-00014 Helsinki University, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Riikka","family":"Keto","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, FIN-00014 Helsinki University, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annukka","family":"Markkula","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, FIN-00014 Helsinki University, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mari","family":"Nevas","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, FIN-00014 Helsinki University, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sebastian","family":"Hielm","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, FIN-00014 Helsinki University, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hannu","family":"Korkeala","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, FIN-00014 Helsinki University, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1472-765X.1997.00204.x"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)38824-6"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X-64.2.201"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1472-765X.1998.00420.x"},{"key":"e_1_3_2_6_2","unstructured":"Buchanan R. 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