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• W2156827123 abstract "Macrolide, lincosamide and streptogramin (MLS) antibiotics are chemically distinct, but are alike in their mode of action, which is inhibition of protein synthesis by binding to the 23S component of the 50S ribosomal subunit. Three mechanisms of MLS resistance are most frequently encountered: (1) enzymatic target modification by an N6 -dimethylase [1Weisblum B Inducible resistance to macrolides, lincosamides and streptogramin type B antibiotics: the resistance phenotype, its biological diversity, and structural elements that regulate expression—a review.J Antimicrob Chemother. 1985; 16 (suppl. A): 63-90Crossref PubMed Google Scholar, 2Lai CJ Weisblum B Altered methylation of ribosomal RNA in an erythromycin-resistant strain of Staphylococcus aureus.Proc Natl Acad Sci USA. 1971; 68: 856-860Crossref PubMed Scopus (190) Google Scholar, 3Duval J Evolution and epidemiology of MLS resistance.J Antimicrob Chemother. 1985; 16 (suppl. A): 137-149Crossref PubMed Google Scholar]; (2) enzymatic inactivation by (a) 3-lin 4-cli O -nucleotidyltransferase (linA), (b) streptogramin A acetyltransferase (saa), and (c) streptogramin B hydrolase (sbh) [4Leclercq R Carlier C Duval J Courvalin P Plasmid-mediated resistance to lincomycin by inactivation in Staphylococcus haemolyticus.Antimicrob Agents Chemother. 1985; 28: 421-424Crossref PubMed Scopus (30) Google Scholar, 5Le Goffic F Capmau ML Bonnet D et al.Plasmid-mediated pristinamycin resistance. PAC-IIA: a new enzyme which modifies pristinamycin IIA.J Antibiot. 1977; 30: 665-669Crossref PubMed Scopus (34) Google Scholar, 6Le Goffic F Capmau ML Abbe J Cerceau C Dublanchet A Duval J Plasmid-mediated pristinamycin resistance: PH-1A, a pristinamycin 1A hydrolase.Ann Microbiol (Paris). 1977; 128: 471-474Google Scholar]; and (3) active efflux resulting in hampered erythromycin permeability (erpA) or macrolide streptogramin permeability (msrA) [7Goldman RC Capobianco JO Role of an energy-dependent efflux pump in plasmid pNE24-mediated resistance to 14- and 15-membered macrolides in Staphylococcus epidermidis.Antimicrob Agents Chemother. 1990; 34: 1973-1980Crossref PubMed Scopus (46) Google Scholar, 8Ross JI Eady EA Cove JH Cunliffe WJ Baumberg S Wootton JC Inducible erythromycin resistance in staphylococci is encoded by a member of the ATP-binding transport super-gene family.Mol Microbiol. 1990; 4: 1207-1214Crossref PubMed Scopus (298) Google Scholar, 9Lampson BC Von David W Parisi JT Novel mechanism for plasmid-mediated erythromycin resistance by pNE24 from Staphylococcus epidermidis.Antimicrob Agents Chemother. 1986; 30: 653-658Crossref PubMed Scopus (29) Google Scholar]. In staphylococci, N6 -dimethylase activity can either be constitutively expressed, or be induced by the presence of a macrolide. Although in the former situation (constitutive expression) resistance to all macrolide, lincosamide and streptogramin B antibiotics is readily detected by agar diffusion, isolates resistant to 14- and 15-membered macrolides may still appear susceptible to clindamycin in the latter (inducible expression) [10Leclercq R Courvalin P Bacterial resistance to macrolide, lincosamide, and streptogramin antibiotics by target modification.Antimicrob Agents Chemother. 1991; 35: 1267-1272Crossref PubMed Scopus (535) Google Scholar]. The National Committee for Clinical Laboratory Standards (NCCLS) guidelines for performing susceptibility testing by disk diffusion do not specifically address this issue [11National Committee for Clinical Laboratory Standards.Performance standards for antimicrobial disc susceptibility tests.Approved standard. 5th edn. NCCLS, Villanova, PA1993Google Scholar], and have been shown to be unreliable in detecting inducible N6-dimethylase activity [12Leclercq R Courvalin P Intrinsic and unusual resistance to macrolide, lincosamide, and streptogramin antibiotics in bacteria.Antimicrob Agents Chemother. 1991; 35: 1273-1276Crossref PubMed Scopus (189) Google Scholar]. To overcome this problem, a modification with the addition of lincomycin has recently been suggested [13Leclercq R Brisson-Noël A Duval J Courvalin P Phenotypic expression and genetic heterogeneity of lincosamide inactivation in Staphylococcus spp.Antimicrob Agents Chemother. 1987; 31: 1887-1891Crossref PubMed Scopus (36) Google Scholar]. During a 3-month period our laboratory augmented the standard susceptibility testing procedure (NCCLS; erythromycin and clindamycin disks not apposed) by including a lincomycin disk on a separate agar plate (Mueller-Hinton 2 cation-adjusted agar, bioMérieux, Crapone, France). Erythromycin (15 μg; Becton Dickinson, Meylan Cedex, France) and clindamycin (2 μg; Becton Dickinson, Meylan Cedex, France) zone diameters were interpreted according to the NCCLS guidelines [11National Committee for Clinical Laboratory Standards.Performance standards for antimicrobial disc susceptibility tests.Approved standard. 5th edn. NCCLS, Villanova, PA1993Google Scholar]. Interpretations of lincomycin (15 μg; bioMérieux, Marcy l'Etoile, France) zone diameters were those provided by the manufacturer (susceptible, ≥ 21 mm; intermediate, 18-20 mm; resistant, ≤ 17 mm). Staphylococci were defined as catalase-positive, Gram-positive cocci in clusters. Staphylococcus aureus and coagulase-negative staphylococci (CNS) were differentiated on the basis of DNase activity and free coagulase production. Doubtful results were confirmed by Staphylococcus aureus Accuprobe (Gen-Probe, San Diego, CA). CNS were not further identified to the species level. In total, 444 consecutive patient isolates (324 S. aureus, and 120 CNS) that were considered clinically significant were investigated. The augmented screening procedure detected 59 (13.3%) isolates revealing reduced susceptibility to any of the three antibiotics comprising 18 (5.5%) S. aureus and 41 (34.2%) CNS, with the following susceptibility patterns: erythromycin, lincomycin and clindamycin resistant, n= 27; erythromycin monoresistant, n = 24; lincomycin monoresistant, n =2; intermediate to lincomycin, n= 1; lincomycin resistant and intermediate to clindamycin, n = 5. Thus, three isolates (S. aureus, n= 1; CNS, n=2) showing reduced susceptibility to lincomycin were only detected by the augmented screening. Thirty-eight of the 59 resistant isolates (erythromycin, lincomycin, and clindamycin resistant, n =19; erythromycin monoresistant, n = 14; lincomycin monoresistant, n =1; intermediate to lincomycin, n =1; lincomycin resistant and intermediate to clindamycin, n =3) were available for retesting both by the NCCLS standard procedure and by a modified disk diffusion method in which the erythromycin disk was placed in between the clindamycin and lincomycin disks (distance 15 mm). From susceptibility patterns (Figure 1) the isolates were classified according to the putative resistance mechanisms [12Leclercq R Courvalin P Intrinsic and unusual resistance to macrolide, lincosamide, and streptogramin antibiotics in bacteria.Antimicrob Agents Chemother. 1991; 35: 1273-1276Crossref PubMed Scopus (189) Google Scholar]: (a) constitutive N6 - dimethylase, n =18 (S. aureus, n =3; CNS, n =15); (b) inducible N6 -dimethylase, n =14 (S. aureus, n= 3; CNS, n =11); (c) linA, n =4 (S. aureus, n =3; CNS, n =1); (d) msrA or erpA, n =1 (CNS); (e) saa or sbh, n =1 (CNS). Differentiating saa from sbh and erpA from msrA was not possible, since streptogramins were not tested [12Leclercq R Courvalin P Intrinsic and unusual resistance to macrolide, lincosamide, and streptogramin antibiotics in bacteria.Antimicrob Agents Chemother. 1991; 35: 1273-1276Crossref PubMed Scopus (189) Google Scholar]. Comparison of the modified to the NCCLS standard procedure revealed unequivocal results regarding the 18 isolates harboring constitutively expressed N6 -dimethylase and one isolate showing msrA or erpA activity. The modified procedure identified inducible N6 -dimethylase activity in 14 additional isolates, irrespective of the lincosamide tested (Figure 1b); linA and saa or sbh activity, respectively, in one isolate each, however, were only detected by lincomycin (Figure 1c,e). By the NCCLS standard procedure (erythromycin and clindamycin disks not apposed), 13 of 14 isolates with inducible N6 -dimethylase activity appeared to be clindamycin-susceptible (Figure 2a), but apposing clindamycin next to erythromycin disks resulted in the detection of lincosamide resistance conferred by this mechanism in all 14 isolates (Figure 2b). Although there is still controversy over the clinical significance of clindamycin resistance conferred by inducible N6 -dimethylase activity, relapse in patients with staphylococcal infections who were treated with clindamycin has been reported [14Watanakunakorn C Clindamycin therapy of Staphylococcus aureus endocarditis. Clinical relapse and development of resistance to clindamycin, lincomycin and erythromycin.Am J Med. 1976; 60: 419-425Abstract Full Text PDF PubMed Scopus (77) Google Scholar]. While it reliably detected inducible N6 -dimethylase activity when apposed next to the erythromycin disk, clindamycin alone detected only three of four isolates with resistance conferred by linA (Figure 1c). The proper detection of this resistance mechanism, which greatly impairs the activities of both lincomycin and clindamycin in vitro [13Leclercq R Brisson-Noël A Duval J Courvalin P Phenotypic expression and genetic heterogeneity of lincosamide inactivation in Staphylococcus spp.Antimicrob Agents Chemother. 1987; 31: 1887-1891Crossref PubMed Scopus (36) Google Scholar], is a prerequisite for assessing its clinical significance, and, since clindamycin is still an important drug for the treatment of staphylococcal infections, the detection of any mechanism conferring resistance to this drug may be important for predicting the clinical outcome. Our findings confirm that the reliable detection of MLS resistance conferred by inducible N6 -dimethylase activity in staphylococci by disk diffusion requires the apposition of the lincosamide next to the erythromycin disk; the substitution of lincomycin for clindamycin further increases the sensitivity of the resistance screening." @default.
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• W2156827123 title "Phenotypes of resistance to macrolide and lincosamide antibiotics in Staphylococcus species" @default.
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