Prevalence and association of PCR ribotypes of Clostridium difficile isolated from symptomatic patients from Warsaw with macrolide-lincosamide-streptogramin B (MLSB) type resistance

Abstract

Isolates (79 in total) of Clostridium difficile obtained over a 2 year period from 785 patients suspected of having C. difficile-associated diarrhoea (CDAD) and being hospitalized in the University Hospital in Warsaw were characterized by toxigenicity profile and PCR ribotyping. Furthermore, their susceptibility to clindamycin and erythromycin was determined. ) to clindamycin and erythromycin was found in 39 (49 %) of the C. difficile isolates. Interestingly, 34 (94 %) of macrolide-lincosamide-streptogramin B (MLS B ) type resistance strains did not produce toxin A, but produced toxin B and were A " B + ribotype 017. Thirty-seven of the high-level resistance strains harboured the erythromycin-resistance methylase gene (ermB). C. difficile isolates (2/29) that had high-level clindamycin and erythromycin resistance, and belonged to PCR ribotype 046, were ermB negative. These investigations revealed that the predominant C. difficile strain isolated from symptomatic patients hospitalized in University Hospital in Warsaw was MLS B -positive clindamycin/erythromycin-resistant PCR ribotype 017.

Characterization and antimicrobial susceptibility of Clostridium difficile strains isolated from adult patients with diarrhoea hospitalized in two university hospitals in Poland, 2004–2006

“…Resistance against clindamycin and erythromycin among Polish A 2 B + CDT 2 (PCR ribotype 017) C. difficile strains was very high (91 %) but not among A + B + CDT 2 and A 2 B 2 CDT 2 strains (7 and 2.1 %, respectively), which confirmed our previous observations (Pituch et al, 2006(Pituch et al, , 2007. Ilchmann et al (2010), in a study performed in Germany, documented a significant increase from 13.0 to 54.8 % of erythromycin-and clindamycin-resistant C. difficile strains belonging to PCR ribotype 001 (Ilchmann et al, 2010).…”

“…Among the strains investigated in that study, 44.3 % were A + B + CDT 2 , 45.5 % were A 2 B + CDT 2 , 1.3 % were A + B + CDT + and 8.9 % were A 2 B 2 CDT 2 (Pituch et al, 2006). During the 2-year study period, one outbreak of CDI cases caused by a C. difficile PCR ribotype 017 strain occurred among 12 patients at the internal unit.…”

“…In a previous study conducted between 2002 and 2003 in Poland, we analysed 79 C. difficile strains isolated over a 2-year period from 785 adult patients with antibioticassociated diarrhoea hospitalized in one university hospital in Warsaw (Pituch et al, 2006). Among the strains investigated in that study, 44.3 % were A + B + CDT 2 , 45.5 % were A 2 B + CDT 2 , 1.3 % were A + B + CDT + and 8.9 % were A 2 B 2 CDT 2 (Pituch et al, 2006).…”

“…PCRs to detect the tcdA and tcdB genes and deletions in the tcdA gene were conducted as described previously (Pituch et al, 2006). Primers described by Stubbs et al (2000) were used for amplification of the CDT genes cdtA and cdtB, as described previously (Pituch et al, 2006).…”

“…Quality-control strains (Bacteroides fragilis NCTC 11295, Bacteroides thetaiotaomicron ATCC 29741, Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923) were always included. A 688 bp fragment of the ermB gene for erythromycin resistance was amplified using specific primer pairs 2980 (59-AATAAGTAAACAGGTAACGTT-39) and 2981 (59-GCTCCTTGGAAGCTGTCAGTAG-39) (Johnson et al, 1999 (Pituch et al, 2006) generated products of 630 and 399 bp for the tcdA and tcdB genes, respectively. PCR to detect deletion of repeat sequences in the tcdA gene with the NK9/ NKV011 (Pituch et al, 2006) primer pair for the 144 A 2 B + CDT 2 strains generated a 700 bp product similar to that obtained for the Japanese GAI 95 601 C. difficile strain and for the prevalent group of A 2 B + strains of PCR ribotype 017.…”