title: Comparison of two RT-PCR methods for quantifying ampC specific transcripts in Escherichia coli strains.
abstract: In Escherichia coli, beta-lactam resistance usually depends on beta-lactamase production. AmpC chromosomal cephalosporinase hyperproduction is generally due to mutations in the ampC gene promoter. In order to study ampC expression in E. coli clinical strains, we have compared two methods: conventional and real-time reverse transcription-polymerase chain reaction (RT-PCR). With both methods, ampC mRNA was found to be greatly increased in strains presenting -42 or -32 mutations in the ampC promoter, and moderately increased when a -11 mutation was present in the Pribnow box. Real-time RT-PCR represents a powerful tool combining amplification, fluorescent detection and analysis.
authors.affiliation: Laboratoire de Bactériologie-Virologie-Hygiène Hospitalière, Institut de Biologie des Hôpitaux de Nantes, 9 quai Moncousu, 44093 Nantes Cedex 01, France. stephane.corvec@chu-nantes.fr
authors.forename: Stéphane
authors.initials: S
authors.lastname: Corvec
authors.forename: Nathalie
authors.initials: N
authors.lastname: Caroff
authors.forename: Eric
authors.initials: E
authors.lastname: Espaze
authors.forename: Julie
authors.initials: J
authors.lastname: Marraillac
authors.forename: Henri
authors.initials: H
authors.lastname: Drugeon
authors.forename: Alain
authors.initials: A
authors.lastname: Reynaud
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D005798:Genes, Bacterial; D008826:Microbial Sensitivity Tests; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014158:Transcription, Genetic; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000235:genetics*; D047090:beta-Lactams / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D047090:beta-Lactams; C048795:AmpC beta-lactamases; D001618:beta-Lactamases