{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T02:42:13Z","timestamp":1792118533094,"version":"4.4.0"},"reference-count":44,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2007,1,1]],"date-time":"2007-01-01T00:00:00Z","timestamp":1167609600000},"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":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2007,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            CTX-M extended-spectrum \u03b2-lactamases (ESBLs) are increasingly prevalent worldwide among\n            <jats:italic>Escherichia coli<\/jats:italic>\n            bacteria, mostly in community-acquired urinary tract infections. Finding a fast and reliable technique for identification of CTX-M enzymes is becoming a challenge for the microbiology laboratory. A fast real-time PCR amplification technique, using degenerated primers specific for all the\n            <jats:italic>bla<\/jats:italic>\n            <jats:sub>CTX-M<\/jats:sub>\n            alleles, coupled to real-time pyrosequencing was developed. The five CTX-M groups were unambiguously identified by pyrosequencing a 13-bp DNA region. Further sequencing of an additional 16-bp region allowed further division into subgroups. Phylogenetic trees constructed with the entire\n            <jats:italic>bla<\/jats:italic>\n            <jats:sub>CTX-M<\/jats:sub>\n            genes and with both pyrosequenced regions (29 bp) gave similar results, suggesting that this technique, termed the real-time detection and sequencing method, has a powerful discriminatory ability. This high-throughput technique has been evaluated by screening 48 ESBL-producing\n            <jats:italic>E. coli<\/jats:italic>\n            isolates recovered from the Bice\u0302tre hospital (France) in 2004. Forty-four of these strains were CTX-M positive by real-time PCR detection and direct pyrosequencing of the PCR products, which identified CTX-M-15 as the main CTX-M-type \u03b2-lactamase. Pulsed-field gel electrophoresis analysis of these strains revealed that several clones, of which one CTX-M-15-positive clone was predominant (60%), were identified both in nosocomial and in community-acquired isolates. The combination of real-time PCR with pyrosequencing represents a powerful tool for epidemiological studies of CTX-M producers. This assay has the potential to be used in a diagnostic laboratory since up to 96 bacterial isolates may be screened in less than 3 h.\n          <\/jats:p>","DOI":"10.1128\/aac.00611-06","type":"journal-article","created":{"date-parts":[[2006,12,20]],"date-time":"2006-12-20T18:32:51Z","timestamp":1166639571000},"page":"223-230","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":83,"title":["Identification of CTX-M-Type Extended-Spectrum-\u03b2-Lactamase Genes Using Real-Time PCR and Pyrosequencing"],"prefix":"10.1128","volume":"51","author":[{"given":"Thierry","family":"Naas","sequence":"first","affiliation":[{"name":"Service de Bacte\u0301riologie-Virologie, Ho\u0302pital de Bice\u0302tre, Assistance Publique\/Ho\u0302pitaux de Paris, Faculte\u0301 de Me\u0301decine Paris-Sud, Universite\u0301 Paris XI, 94275 Le Kremlin-Bice\u0302tre, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cynthia","family":"Oxacelay","sequence":"additional","affiliation":[{"name":"Service de Bacte\u0301riologie-Virologie, Ho\u0302pital de Bice\u0302tre, Assistance Publique\/Ho\u0302pitaux de Paris, Faculte\u0301 de Me\u0301decine Paris-Sud, Universite\u0301 Paris XI, 94275 Le Kremlin-Bice\u0302tre, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrice","family":"Nordmann","sequence":"additional","affiliation":[{"name":"Service de Bacte\u0301riologie-Virologie, Ho\u0302pital de Bice\u0302tre, Assistance Publique\/Ho\u0302pitaux de Paris, Faculte\u0301 de Me\u0301decine Paris-Sud, Universite\u0301 Paris XI, 94275 Le Kremlin-Bice\u0302tre, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"203","volume":"134","year":"1995","unstructured":"Arlet, G., G. 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