A New Resistance Gene, linB , Conferring Resistance to Lincosamides by Nucleotidylation in Enterococcus faecium HM1025

Abstract

Resistance to lincomycin and clindamycin in the clinical isolateEnterococcus faecium HM1025 is due to a ribosomal methylase encoded by an ermAM-like gene and the plasmid-mediated inactivation of these antibiotics. We have cloned and determined the nucleotide sequence of the gene responsible for the inactivation of lincosamides, linB. This gene encodes a 267-amino-acid lincosamide nucleotidyltransferase. The enzyme catalyzes 3-(5′-adenylation) (the adenylation of the hydroxyl group in position 3 of the molecules) of lincomycin and clindamycin. Expression oflinB was observed in both Escherichia coli andStaphylococcus aureus. The deduced amino acid sequence of the enzyme did not display any significant homology with staphylococcal nucleotidyltransferases encoded by linA andlinA′ genes. Sequences homologous to linB were found in 14 other clinical isolates of E. faecium, indicating the spread of the resistance trait in this species.

Susceptibility ofClostridium perfringensstrains from broiler chickens to antibiotics and anticoccidials

“…The PCR running conditions consisted of an initial cycle of 5 min of denaturation at 948C, followed by 35 cycles of 1 min denaturation at 948C, 1 min of annealing at the annealing temperatures presented in Table 1 and 1 min of elongation at 728C, followed by a final extension of 5 min at 728C. The reference strains Staphylococcus haemolyticus BM4610 (lnu (A)), Enterococcus faecalis JH2-2/1025 (lnu (B)), Escherichia coli EC6 (tet (M)), E. coli EC7 (tet (K)), E. coli C65 (tet (L)) and E. coli EC70 (tet P(B)) were used as positive controls (Abraham & Rood, 1985;Brisson-Noël & Courvalin, 1986;Morse et al , 1986, Schwartz et al , 1992Bozdogan et al , 1999).…”

Molecular basis of resistance to macrolides and lincosamides among staphylococci and streptococci from various animal sources collected in the resistance monitoring program BfT-GermVet

“…Positive controls for the genes erm(A), erm(B), erm(C), msr(A), mph(C) and lnu(A) were derived from a previous study [5]. Positive controls for lnu(B) [8] and lnu(C) [9] were kindly provided by Roland Leclercq. At least one amplicon for msr(D), mef(A) and lnu(B) was cloned and sequenced for control purposes.…”

“…PCR assays for the following genes were carried out according to previously described protocols: erm(A) [13], erm(TR) (subtype of the erm(A) class) [14], erm(B) [15], erm(C) [16], mef(A) [17], msr(A) [18], mph(C) [5], lnu(A) [19], lnu(B) [8] and lnu(C). The amplified region in the erm(A) and erm(C) PCR assays comprises the regulatory region of these genes in order to confirm the inducible or constitutive resistance phenotype.…”

“…The mph(C) gene, coding for macrolide phosphotransferase C (Mph(C)), has recently been detected in low-level resistant staphylococcal isolates of bovine [5] and equine [6] origin. Three classes of lincosamide nucleotidyltransferases (Lnu(A), Lnu(B) and Lnu(C)) are known in Gram-positive bacteria and are encoded by the genes lnu(A) [7], lnu(B) [8] and lnu(C) [9], respectively.…”

Nosocomial outbreak of methicillin- and linezolid-resistant Staphylococcus epidermidis associated with catheter-related infections in intensive care unit patients

“…Furthermore, a PCR was performed to detect the presence of the cfr gene (Kehrenberg and Schwarz, 2006). The presence of genes associated with methicillin [mecA], aminoglycosides [ant(6)-Ia, aac(6 )-Ie-aph(2 )-Ia, aph(3 )-IIIa, and ant(4 )], lincomycin [linA, linB], and chloramphenicol resistance [catA] was analysed by PCR (van de Klundert and Vliegenthart, 1993;Bozdogan et al, 1999;Lina et al, 1999;Aarestrup et al, 2000;Kondo et al, 2007). The presence of the staphylococcal chromosomal cassette mec (SCCmec) type was determined by multiplex PCR assays (Kondo et al, 2007), and the detection of ica operon and IS256 element was performed by PCR (Ziebuhr et al, 1999).…”