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).
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.
Klomberg
Debby M
DM
Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Weg door Jonkerbos 100, 6532 SZ Nijmegen, The Netherlands.
de Valk
Hanneke A
HA
Mouton
Johan W
JW
Klaassen
Corné H W
CH
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*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C117866:MefA protein, Streptococcus; D008565:Membrane Proteins