{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T13:09:31Z","timestamp":1762175371565},"reference-count":26,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2000,4,1]],"date-time":"2000-04-01T00:00:00Z","timestamp":954547200000},"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":[[2000,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:italic>Clostridium botulinum<\/jats:italic>\n            produces a characteristic botulinum neurotoxin which can cause an often fatal neuroparalytic condition known as botulism. Although food-borne botulism is rare, critical screening by food companies is necessary to ensure that food products are safe. At present, the food industry assesses the risks of botulinum neurotoxin production by challenge testing to check any new food products and to check the efficacy of new storage regimes. Challenge testing involves artificial introduction of defined strains of microorganisms into food, and microbial growth and possible toxin production are then monitored. Botulinum toxin is normally analyzed by using the mouse bioassay. However, the mouse bioassay is expensive, slow, and politically sensitive because of animal rights issues. In this paper we describe adaptation of a new assay, competitive reverse transcription-PCR (RT-PCR), to monitor botulinum neurotoxin production. This method accurately measures the level of toxin-encoding mRNA in\n            <jats:italic>C. botulinum<\/jats:italic>\n            cells. Measurement of mRNA should provide a good indication of gene expression as mRNA is turned over rapidly in bacterial cells. In addition, the method is rapid, specific, and sensitive. The competitive RT-PCR method was developed to examine\n            <jats:italic>C. botulinum<\/jats:italic>\n            E VH toxin gene expression and was used to investigate the level of toxin production by\n            <jats:italic>C. botulinum<\/jats:italic>\n            E VH when the organism was grown in two different types of broth. The results which we obtained with the competitive RT-PCR method demonstrated that this method is more rapid and more sensitive than the mouse bioassay.\n          <\/jats:p>","DOI":"10.1128\/aem.66.4.1423-1428.2000","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:56:57Z","timestamp":1027763817000},"page":"1423-1428","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":28,"title":["Quantification of\n            <i>Clostridium botulinum<\/i>\n            Toxin Gene Expression by Competitive Reverse Transcription-PCR"],"prefix":"10.1128","volume":"66","author":[{"given":"S.","family":"McGrath","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->School of Applied Biological and Chemical Science, University of Ulster, Coleraine, Co. Londonderry, Northern Ireland, BT52 1SA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J. S. G.","family":"Dooley","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->School of Applied Biological and Chemical Science, University of Ulster, Coleraine, Co. Londonderry, Northern Ireland, BT52 1SA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. W.","family":"Haylock","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->School of Applied Biological and Chemical Science, University of Ulster, Coleraine, Co. Londonderry, Northern Ireland, BT52 1SA"}],"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":"10.1007\/BF02762407"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.31.9.2255-2262.1993"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.62.8.3069-3072.1996"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2672.1997.00313.x"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0168-9525(93)90137-7"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2275(20)40095-1"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/0963-9969(92)90171-Z"},{"key":"e_1_3_2_10_2","unstructured":"Hatheway\nC. L. Clostridium botulinum and other clostridia that produce botulinum toxin\nClostridium botulinum: ecology and control in foods.\nHauschild\nA. H. W.\nDodds\nK. L.\n1992\n3\n20\nMarcel Dekker Inc.\nNew York N.Y"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(88)61023-8"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.5.1161-1165.1997"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.63.11.4441-4448.1997"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X-56.9.769"},{"key":"e_1_3_2_15_2","doi-asserted-by":"crossref","unstructured":"Middlebrook\nJ. L.\nCell surface receptors for protein toxins\nBotulinum neurotoxin and tetanus toxin.\nSimpson\nL. L.\n1989\n95\n119\nAcademic Press\nSan Diego Calif","DOI":"10.1016\/B978-0-12-644445-2.50011-8"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(82)84020-2"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X-56.10.856"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1006\/mcpr.1998.0184"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/0163-7258(82)90061-4"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.61.11.3981-3985.1995"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.4.1313-1318.1998"},{"key":"e_1_3_2_22_2","first-page":"16","article-title":"Kinetic studies on the interaction between botulinum toxin type A and the cholinergic neuromuscular junction.","volume":"212","author":"Simpson L. L.","year":"1980","unstructured":"Simpson L. L. Kinetic studies on the interaction between botulinum toxin type A and the cholinergic neuromuscular junction. J. Pharmacol. Exp. Ther. 212 1980 16 21","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.2144\/96212rr01"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2672.1994.tb01650.x"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.2144\/96215st04"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2672.1998.0840s187S.x"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/0923-2508(93)90004-L"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.66.4.1423-1428.2000","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.66.4.1423-1428.2000","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T09:44:07Z","timestamp":1645609447000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.66.4.1423-1428.2000"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2000,4]]},"references-count":26,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2000,4]]}},"alternative-id":["10.1128\/AEM.66.4.1423-1428.2000"],"URL":"https:\/\/doi.org\/10.1128\/aem.66.4.1423-1428.2000","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2000,4]]},"assertion":[{"value":"1999-09-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2000-01-07","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2000-04-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}