title: Rapid and reliable real-time PCR assay for detection of the macrolide efflux gene and subsequent discrimination between its distinct subclasses mef(A) and mef(E).
abstract: A real-time PCR assay is described for detection of the macrolide efflux gene, mef. Following amplification, unambiguous discrimination between the two mef subclasses, mef(A) and mef(E), is easily established using a melting curve analysis. The results of this novel assay were 100% concordant with a conventional PCR-RFLP approach but requires far less hands-on time. Furthermore, the real-time format offers semiquantitative results allowing identification of contaminated cultures and/or DNA preparations.
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Weg door Jonkerbos 100, 6532 SZ Nijmegen, The Netherlands.
authors.forename: Debby M
authors.initials: DM
authors.lastname: Klomberg
authors.forename: Hanneke A
authors.initials: HA
authors.lastname: de Valk
authors.forename: Johan W
authors.initials: JW
authors.lastname: Mouton
authors.forename: Corné H W
authors.initials: CH
authors.lastname: Klaassen
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000097:blood / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013296:Streptococcus pneumoniae / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C117866:MefA protein, Streptococcus; D008565:Membrane Proteins