title: Characterisation and molecular cloning of the novel macrolide-streptogramin B resistance determinant from Staphylococcus epidermidis.
abstract: A total of 110 staphylococcal isolates from human skin were found to express a novel type of erythromycin resistance. The bacteria were resistant to 14-membered ring macrolides (MIC 32-128 mg/l) but were sensitive to 16-membered ring macrolides and lincosamides. Resistance to type B streptogramins was inducible by erythromycin. A similar phenotype, designated MS resistance, was previously described in clinical isolates of coagulase-negative staphylococci from the USA. In the UK, MS resistance is widely distributed in coagulase-negative staphylococci but was not detected in 100 erythromycin resistant clinical isolates of Staphylococcus aureus. Tests for susceptibility to a further 16 antibiotics failed to reveal any other selectable marker associated with the MS phenotype. Plasmid pattern analysis of 48 MS isolates showed considerable variability between strains and no common locus for the resistance determinant. In one strain of S. epidermidis co-resistance to tetracycline, penicillin and erythromycin (MS) was associated with a 31.5 kb plasmid, pUL5050 which replicated and expressed all three resistances when transformed into S. aureus RN4220. The MS resistance determinant was localised to a 1.9 kb fragment which was cloned on to the high-copy-number vector, pSK265. A constitutive mutant of S. aureus RN4220 containing the 1.9 kb fragment remained sensitive to clindamycin. This observation, together with the concentration-dependent induction (optimum 5 mg/l of erythromycin) of virginiamycin S resistance suggests that the MS phenotype is not due to altered expression of MLS resistance determinants (erm genes) but probably occurs via a different mechanism.
authors.affiliation: Department of Microbiology, University of Leeds, UK.
authors.forename: J I
authors.initials: JI
authors.lastname: Ross
authors.forename: A M
authors.initials: AM
authors.lastname: Farrell
authors.forename: E A
authors.initials: EA
authors.lastname: Eady
authors.forename: J H
authors.initials: JH
authors.lastname: Cove
authors.forename: W J
authors.initials: WJ
authors.lastname: Cunliffe
mesh_terms: D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D004352:Drug Resistance, Microbial; D008826:Microbial Sensitivity Tests; D010641:Phenotype; D010957:Plasmids; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics; D013212:Staphylococcus epidermidis / Q000187:drug effects / Q000235:genetics*; D014169:Transformation, Bacterial; D014769:Virginiamycin / Q000494:pharmacology*
chemical_list: D004269:DNA, Bacterial; D014769:Virginiamycin; D004262:DNA Restriction Enzymes