{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T14:18:31Z","timestamp":1791296311223,"version":"4.1.0"},"reference-count":24,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2002,4,1]],"date-time":"2002-04-01T00:00:00Z","timestamp":1017619200000},"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":["J Clin Microbiol"],"published-print":{"date-parts":[[2002,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            We have developed a rapid (1-h) real-time fluorescence-based PCR assay with the Smart Cycler thermal cycler (Cepheid, Sunnyvale, Calif.) for the detection of Shiga toxin-producing\n            <jats:italic>Escherichia coli<\/jats:italic>\n            (STEC), as well as other Shiga toxin-producing bacteria. Based on multiple-sequence alignments, we have designed two pairs of PCR primers that efficiently amplify all variants of the Shiga toxin genes\n            <jats:italic>stx<\/jats:italic>\n            <jats:sub>1<\/jats:sub>\n            and\n            <jats:italic>stx<\/jats:italic>\n            <jats:sub>2<\/jats:sub>\n            , respectively. These primer pairs were combined for use in a multiplex assay. Two molecular beacons bearing different fluorophores were used as internal probes specific for each amplicon. Assays performed with purified genomic DNA from a variety of STEC strains (\n            <jats:italic>n<\/jats:italic>\n            = 23) from diverse geographic locations showed analytical sensitivities of about 10 genome copies per PCR. Non-STEC strains (\n            <jats:italic>n<\/jats:italic>\n            = 20) were also tested, and no amplification was observed. The PCR results correlated perfectly with the phenotypic characterization of toxin production in both STEC and non-STEC strains, thereby confirming the specificity of the assay. The assay was validated by testing 38 fecal samples obtained from 27 patients. Of these samples, 26 were PCR positive for\n            <jats:italic>stx<\/jats:italic>\n            <jats:sub>1<\/jats:sub>\n            and\/or\n            <jats:italic>stx<\/jats:italic>\n            <jats:sub>2<\/jats:sub>\n            . Compared with the culture results, both the sensitivity and the negative predictive value were 100%. The specificity was 92%, and the positive predictive value was 96%. Moreover, this assay detected STEC from a sample in which the STEC concentration was at the limit of detection of the conventional culture methods and from a sample in which STEC was not detected by the conventional culture methods. This real-time PCR assay is simple, rapid, sensitive, and specific and allows detection of all Shiga toxin-producing bacteria directly from fecal samples, irrespective of their serotypes.\n          <\/jats:p>","DOI":"10.1128\/jcm.40.4.1436-1440.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:59:40Z","timestamp":1027763980000},"page":"1436-1440","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":80,"title":["Rapid Detection of Shiga Toxin-Producing Bacteria in Feces by Multiplex PCR with Molecular Beacons on the Smart Cycler"],"prefix":"10.1128","volume":"40","author":[{"given":"Simon D.","family":"Be\u0301langer","sequence":"first","affiliation":[{"name":"Centre de Recherche en Infectiologie de l'Universite\u0301 Laval, Centre Hospitalier Universitaire de Que\u0301bec (Pavillon Centre Hospitalier de l'Universite\u0301 Laval)"},{"name":"Division de Microbiologie, Faculte\u0301 de Me\u0301decine, Universite\u0301 Laval, Sainte-Foy, Que\u0301bec, Canada G1V 4G2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maurice","family":"Boissinot","sequence":"additional","affiliation":[{"name":"Centre de Recherche en Infectiologie de l'Universite\u0301 Laval, Centre Hospitalier Universitaire de Que\u0301bec (Pavillon Centre Hospitalier de l'Universite\u0301 Laval)"},{"name":"Division de Microbiologie, Faculte\u0301 de Me\u0301decine, Universite\u0301 Laval, Sainte-Foy, Que\u0301bec, Canada G1V 4G2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Me\u0301nard","sequence":"additional","affiliation":[{"name":"Infectio Diagnostic (I.D.I.), Inc., Sainte-Foy, Que\u0301bec, Canada G1V 2K8"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franc\u0327ois J.","family":"Picard","sequence":"additional","affiliation":[{"name":"Centre de Recherche en Infectiologie de l'Universite\u0301 Laval, Centre Hospitalier Universitaire de Que\u0301bec (Pavillon Centre Hospitalier de l'Universite\u0301 Laval)"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michel G.","family":"Bergeron","sequence":"additional","affiliation":[{"name":"Centre de Recherche en Infectiologie de l'Universite\u0301 Laval, Centre Hospitalier Universitaire de Que\u0301bec (Pavillon Centre Hospitalier de l'Universite\u0301 Laval)"},{"name":"Division de Microbiologie, Faculte\u0301 de Me\u0301decine, Universite\u0301 Laval, Sainte-Foy, Que\u0301bec, Canada G1V 4G2"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0923-2508(98)80001-6"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM200007203430303"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.med.50.1.355"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2672.2000.tb05331.x"},{"key":"e_1_3_2_6_2","first-page":"459","year":"1999","unstructured":"Bopp, C. 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