An investigation of the molecular mechanisms contributing to high-level erythromycin resistance in Campylobacter

Corcoran, Deborah; Quinn, Teresa; Cotter, Lesley; Fanning, Séamus

doi:10.1016/j.ijantimicag.2005.08.019

Cited by 83 publications

(77 citation statements)

References 28 publications

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“…A number of previous studies have reported modifications in the ribosomal proteins L4 and L22 (3,4,6,19). Several amino acid substitutions in the ribosomal protein L22 were observed also in the present study.…”

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confidence: 86%

“…The most important macrolide resistance mechanisms in Campylobacter are due to modifications of the ribosomal target sites. High-level resistance is mainly caused by mutations at positions 2058 and 2059 (Escherichia coli numbering) of the 23S rRNA gene (6,7,17). Campylobacters contain three copies of this gene (8).…”

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confidence: 99%

“…Campylobacters contain three copies of this gene (8). Several modifications in the ribosomal proteins L4 and L22, associated with macrolide resistance in other bacteria (5, 13, 15), have been reported in Campylobacter (3,4,6,19). We used pyrosequencing to examine the role of 23S rRNA mutations as a cause of macrolide resistance in a large collection of Campylobacter strains.…”

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confidence: 99%

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Ribosomal Mutations as the Main Cause of Macrolide Resistance in Campylobacter jejuni and Campylobacter coli

Lehtopolku

Kotilainen

Haanperä-Heikkinen³

et al. 2011

Antimicrob Agents Chemother

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The aim of this study was to examine macrolide resistance mutations in Campylobacter species. In 76 strains studied, point mutation A to G at position 2059 of the 23S rRNA gene was detected in 30 of the 33 erythromycinresistant strains. An amino acid insertion in the ribosomal protein L22 was found in one resistant strain without a 23S rRNA mutation. The A2059G mutation is the main cause of macrolide resistance in Campylobacter species.Campylobacters are recognized as a leading cause of bacterial gastroenteritis worldwide (1). The most important macrolide resistance mechanisms in Campylobacter are due to modifications of the ribosomal target sites. High-level resistance is mainly caused by mutations at positions 2058 and 2059 (Escherichia coli numbering) of the 23S rRNA gene (6,7,17). Campylobacters contain three copies of this gene (8). Several modifications in the ribosomal proteins L4 and L22, associated with macrolide resistance in other bacteria (5, 13, 15), have been reported in Campylobacter (3,4,6,19). We used pyrosequencing to examine the role of 23S rRNA mutations as a cause of macrolide resistance in a large collection of Campylobacter strains. We also sequenced the L4 and L22 ribosomal protein genes as well as evaluating the effect of the efflux pump inhibitors on the MIC values.(This work was presented in part at the 14th International Workshop on Campylobacter, Helicobacter and Related Organisms, Rotterdam, The Netherlands, 2007 [P351].)The study collection consisted of 76 Campylobacter strains isolated from stool specimens of Finnish patients between 2003 and 2008. Of these strains, 53 were from an earlier study collection (14), and 23 strains were collected locally in the area of the Turku University Hospital. The cultivation of the stool samples, the preliminary identification of the isolates, and MIC determinations were done as described previously (14). The MIC breakpoint for erythromycin was Ն16 g/ml (11). The hippurate hydrolysis test was used for final species identification (16). Campylobacter jejuni DSM 4688 type strain and Campylobacter coli DSM 4689 type strain (both erythromycin susceptible with wild-type 23S rRNA) were used as control strains. Mutations at positions 2058 and 2059 (E. coli numbering) of the 23S rRNA gene were analyzed by pyrosequencing as described by Haanperä et al. (10). L4 and L22 gene sequences of all 33 erythromycin-resistant and 20 erythromycinsusceptible Campylobacter strains were determined using standard Sanger sequencing using primers 5Ј-GAATTTGCTCCA ACACGC-3Ј and 5Ј-ACCATCTTGATTCCCAGTTTC-3Ј for L22 protein and primers 5Ј-GTAGTTAAAGGTGCAGTACC

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confidence: 86%

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confidence: 99%

mentioning

confidence: 99%

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Ribosomal Mutations as the Main Cause of Macrolide Resistance in Campylobacter jejuni and Campylobacter coli

Lehtopolku

Kotilainen

Haanperä-Heikkinen³

et al. 2011

Antimicrob Agents Chemother

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“…Similar mutations are responsible for resistance in Helicobacter pylori (20). While different mutations occur at comparable frequencies in Helicobacter, the change of A to G at position 2059 (Escherichia coli equivalent; corresponds to position 2075 of Campylobacter) dominates macrolide resistance in Campylobacter (1,4,5,20,23). Rarely, efflux pumps are responsible for a low level of resistance that is of limited clinical relevance at standard antibiotic doses (3,5,12).…”

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confidence: 99%

“…The theoretical applicability of these probes for Campylobacter was checked by consulting previous publications (1,4,5,6,7,12,17,23) and by using the publicly available ARB program (www.ARB-home.de). The wild-type probe (C wt 23S [CGG GGT CTT TCC GTC TT]) was labeled with 6-carboxyfluorescein (6-FAM) and used in combination with a Cy3-labeled resistance probe (C res 23S 2059AϾG [CGG GGT CTC TCC GTC TT]) at a formamide concentration of 40% in the hybridization buffer and corresponding salt concentrations in the wash buffer as previously described for H. pylori (Thermo Hybaid, Ulm, Germany) (20,22,23).…”

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confidence: 99%