{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T19:53:02Z","timestamp":1789847582479,"version":"4.0.1"},"reference-count":59,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2011,1,1]],"date-time":"2011-01-01T00:00:00Z","timestamp":1293840000000},"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":[[2011,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Due to the severity of the food-borne infection listeriosis, strict legislation governs the detectable and permissible limits at which<jats:italic>Listeria monocytogenes<\/jats:italic>is permitted in foods. These requirements, coupled with the ubiquitous nature of<jats:italic>L. monocytogenes<\/jats:italic>strains and the potential for epidemic outbreaks, mean that the pathogen can devastate affected sectors of the food industry. Although almost all<jats:italic>L. monocytogenes<\/jats:italic>strains have the potential to cause listeriosis, those implicated in the vast majority of epidemics belong to a subset of strains belonging to evolutionary lineage I. It has been established that a significant proportion of these strains, including those implicated in the majority of outbreaks, produce an additional hemolysin, designated listeriolysin S (LLS), which may be responsible for the enhanced virulence of these strains. In order to ultimately establish this definitively, it is important to first be able to rapidly discriminate between LLS-positive and -negative strains. Here, after essential genes within the LLS-encoding cluster,<jats:italic>Listeria<\/jats:italic>pathogenicity island 3, were identified by deletion mutagenesis, a real-time PCR assay which targets one such gene,<jats:italic>llsX<\/jats:italic>, was developed as a means of identifying LLS-positive<jats:italic>L. monocytogenes<\/jats:italic>. The specificity of the assay was validated against a panel of 40<jats:italic>L. monocytogenes<\/jats:italic>strains (20 of which were LLS positive) and 25 strains representative of other<jats:italic>Listeria<\/jats:italic>species. Furthermore, 1 CFU of an LLS-positive strain per 25 g\/ml of spiked foods was detected in less than 30 h when the assay was coupled with culture enrichment. The detection limit of this assay was 10 genome equivalents.<\/jats:p>","DOI":"10.1128\/aem.01673-10","type":"journal-article","created":{"date-parts":[[2010,11,13]],"date-time":"2010-11-13T05:59:41Z","timestamp":1289627981000},"page":"163-171","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":64,"title":["Real-Time PCR Assay To Differentiate Listeriolysin S-Positive and -Negative Strains of<i>Listeria monocytogenes<\/i>"],"prefix":"10.1128","volume":"77","author":[{"given":"Evelyn M.","family":"Clayton","sequence":"first","affiliation":[{"name":"Department of Microbiology, University College Cork, Cork, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colin","family":"Hill","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University College Cork, Cork, Ireland"},{"name":"Alimentary Pharmabiotic Centre, Cork, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul D.","family":"Cotter","sequence":"additional","affiliation":[{"name":"Alimentary Pharmabiotic Centre, Cork, Ireland"},{"name":"Teagasc, Moorepark Food Research Centre, Fermoy, County Cork, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. Paul","family":"Ross","sequence":"additional","affiliation":[{"name":"Alimentary Pharmabiotic Centre, Cork, Ireland"},{"name":"Teagasc, Moorepark Food Research Centre, Fermoy, County Cork, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-0691.2009.03109.x"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/25.17.3389"},{"key":"e_1_3_2_4_2","first-page":"1097","volume":"57","year":"2008","unstructured":"Anonymous. 2008. Outbreak of Listeria monocytogenes infections associated with pasteurized milk from a local dairy\u2014Massachusetts, 2007. Morb. Mortal. Wkly. Rep. 57:1097-1100.","journal-title":"Morb. Mortal. Wkly. Rep."},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.fm.2009.07.003"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1086\/591131"},{"key":"e_1_3_2_7_2","first-page":"507","volume":"189","year":"2005","unstructured":"Berche, P. 2005. Pathophysiology and epidemiology of listeriosis. Bull. Acad. Natl. Med. 189:507-516; discussion, 516-521.","journal-title":"Bull. Acad. Natl. Med."},{"key":"e_1_3_2_8_2","first-page":"5","volume":"1","year":"1999","unstructured":"Caplin, B. E., R. P. Rasmussen, P. S. Bernard, and C. T. Wittwer. 1999. The most direct way to monitor PCR amplification for quantification and mutation detection. Biochemica 1:5-8.","journal-title":"Biochemica"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000144"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro932"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2005.04583.x"},{"key":"e_1_3_2_12_2","first-page":"8082","volume":"13","year":"2008","unstructured":"Denny, J., and J. McLauchlin. 2008. Human Listeria monocytogenes infections in Europe: an opportunity for improved European surveillance. Euro Surveill. 13:8082.","journal-title":"Euro Surveill."},{"key":"e_1_3_2_13_2","first-page":"183","volume":"59","year":"2001","unstructured":"Donnelly, C. W. 2001. Listeria monocytogenes: a continuing challenge. Nutr. Rev. 59:183-194.","journal-title":"Nutr. Rev."},{"key":"e_1_3_2_14_2","first-page":"1","volume":"338","year":"2005","unstructured":"European Commission. 2005. Commission regulation (EC) no. 2073\/2005 of 15 November 2005 on microbiological criteria for foodstuffs. Off. J. Eur. Union L338:1-26.","journal-title":"J. Eur. Union"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02912-06"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2171"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.femsre.2004.12.002"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.fm.2009.10.009"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.10.5833-5841.2004"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1413"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2007.05.003"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1995.tb02295.x"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(87)90131-4"},{"key":"e_1_3_2_24_2","first-page":"528","volume":"8","year":"1990","unstructured":"Horton, R. M., Z. L. Cai, S. N. Ho, and L. R. Pease. 1990. Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. Biotechniques 8:528-535.","journal-title":"Biotechniques"},{"key":"e_1_3_2_25_2","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1080\/10408690490489378","volume":"44","year":"2004","unstructured":"Ivanek, R., Y. T. Grohn, L. W. Tauer, and M. Wiedmann. 2004. The cost and benefit of Listeria monocytogenes food safety measures. Crit. Rev. Food Sci. Nutr. 44:513-523.","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268810000294"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-147-5-1095"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.20506\/rst.25.2.1681"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.7.3420-3422.1998"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00508-008-1130-2"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X-65.11.1811"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1093\/qjmed\/hcp181"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01553-09"},{"key":"e_1_3_2_34_2","first-page":"85","volume":"11","year":"2006","unstructured":"Koch, J., and K. Stark. 2006. Significant increase of listeriosis in Germany: epidemiological patterns 2001-2005. Euro Surveill. 11:85-88.","journal-title":"Euro Surveill."},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5290.1188"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.1089\/fpd.2009.0439"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2007.11.066"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1090-0233(97)80005-6"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00474-07"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01963840"},{"key":"e_1_3_2_41_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M900802200"},{"key":"e_1_3_2_42_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00997-08"},{"key":"e_1_3_2_43_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.71.12.8764-8772.2005"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.7.4245-4254.2000"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.fm.2007.07.007"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.fm.2008.08.009"},{"key":"e_1_3_2_47_2","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-148-6-1855"},{"key":"e_1_3_2_48_2","doi-asserted-by":"publisher","DOI":"10.1292\/jvms.09-0247"},{"key":"e_1_3_2_49_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-003-2225-6"},{"key":"e_1_3_2_50_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.3.1366-1377.2004"},{"key":"e_1_3_2_51_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X-68.7.1467"},{"key":"e_1_3_2_52_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.10.6299-6301.2004"},{"key":"e_1_3_2_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.resmic.2006.03.003"},{"key":"e_1_3_2_54_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2007.05.011"},{"key":"e_1_3_2_55_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1605(00)00332-9"},{"key":"e_1_3_2_56_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2009.05.002"},{"key":"e_1_3_2_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01138-08"},{"key":"e_1_3_2_58_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.14.3.584-640.2001"},{"key":"e_1_3_2_59_2","doi-asserted-by":"publisher","DOI":"10.1086\/430625"},{"key":"e_1_3_2_60_2","first-page":"19482","volume":"15","year":"2010","unstructured":"Yde, M., N. Botteldoorn, S. Bertrand, J. Collard, and K. Dierick. 2010. Microbiological and molecular investigation of an increase of human listeriosis in Belgium, 2006-2007. Euro Surveill. 15:19482.","journal-title":"Euro Surveill."}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.01673-10","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.01673-10","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,1]],"date-time":"2024-04-01T06:11:42Z","timestamp":1711951902000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.01673-10"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,1]]},"references-count":59,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2011,1]]}},"alternative-id":["10.1128\/AEM.01673-10"],"URL":"https:\/\/doi.org\/10.1128\/aem.01673-10","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,1]]},"assertion":[{"value":"2010-07-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2010-10-29","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2011-12-24","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}