PCR-RFLP detection of point mutations A2143G and A2142G in 23S rRNA gene conferring resistance to clarithromycin in Helicobacter pylori strains

Background. The occurrence of clarithromycin resistance among Helicobacter pylori strains is a major cause of the treatment failure. Resistance to this drug is conferred by point mutations in 23S rRNA gene and the most prevalent mutations are A2143G and A2142G. The aim of the study was to evaluate the occurrence of A2143G and A2142G mutations in a group of H. pylori strains resistant to clarithromycin. Materials and Methods. The study included 21 clarithromycin-resistant H. pylori strains collected between 2006 and 2009 in southern Poland. Resistance to clarithromycin was quantitatively tested with the E-test to determine the minimal inhibitory concentration (MIC value). The point mutations of H. pylori isolates were detected by PCR followed by RFLP analysis. Results. The MIC values for clarithromycin for the analyzed strains ranged from 1.5 mg/L to 64 mg/L. Nine H. pylori strains exhibited A2143G mutation and A2142G mutation was found in 9 isolates as well. The results of RFLP analysis of 3 clarithromycin-resistant strains were negative for both mutations. The average MIC values for A2143G and A2142G mutants were 6 and 30 mg/L, respectively. Conclusions. Frequencies of A2143G and A2142G mutations were the same in all isolates tested. Strains with A2143G mutation exhibited lower MIC values than A2142G mutants. Application of PCR-RFLP method for detection of clarithromycin resistance allows for better and more efficient management of H. pylori infections

Variability in Prevalence of Helicobacter pylori Strains Resistant to Clarithromycin and Levofloxacin in Southern Poland

Background. An increasing resistance of Helicobacter pylori strains to antimicrobial agents is the serious therapeutic problem. The aim of this study was to compare the primary and secondary resistance of H. pylori strains isolated between 2006–2008 (data published) and 2009–2011 to clarithromycin and levofloxacin. Material and Methods. 220 dyspeptic patients (153 before treatment, 67 after), were enrolled in the study. 51 H. pylori strains were isolated. MIC values of clarithromycin and levofloxacin were determined by the E-test method. The statistical analysis was conducted with the χ2 test with Yates correction at the 0.05 significance level (P ≤ 0.05). Results. Between 2006 and 2008, 34% (39/115) of H. pylori strains were resistant to clarithromycin (primary 21% (19/90), secondary 80% (20/25)). 5% (6/115) of strains were resistant to levofloxacin (primary 2% (2/90), secondary 16% ((4/25); data published) Between 2009–2011, 22% (11/51) of H. pylori strains were resistant to clarithromycin (primary 19% (8/43), secondary 38% (3/8)). 16% (8/51) of strains were resistant to levofloxacin (primary 12% (5/43), secondary 38% (3/8)). Conclusion. The present study has shown the increasing amount of resistant H. pylori strains isolated from patients in Southern Poland to levofloxacin and decreasing number of resistant strains to clarithromycin

Levofloxacin resistance of "Helicobacter pylori" strains isolated from patients in Southern Poland between 2006-2012

An increasing resistance of Helicobacter pylori (H. pylori) to antimicrobial agents leads to the need of regional monitoring of the prevalence resistant strains (according to the Maastricht/Florence consensus report, 2012). The aim of the study was to assess the resistance to levofloxacin of H. pylori strains isolated from adult patients of Ma≥opolska region in Poland. Bioptates taken from gastric mucosa during gastroscopy constituted the material for the study. Two hundred ten H. pylori strains were isolated from 811 patients. A majority of strains (171) came from patients before the treatment of H. pylori infections while the remaining 39 strains were isolated from patients after the failed therapy. Susceptibility of H. pylori to levofloxacin was determined by strips impregnated with antibiotic gradient (E-test, bioMerieux). The obtained minimum inhibitory concentration (MIC) values ranged from 0.002 mg/L to 32 mg/L. The percentage of strains resistant to levofloxacin amounted to 8.10% (17/210). Among the group of strains isolated from patients before the treatment, 5.85% (10/171) of H. pylori strains were resistant to levofloxacin. In the group of strains isolated from patients after the treatment 17.95% (7/39) of strains were resistant. The difference in the frequency of H. pylori strains resistant to levofloxacin in patients before and after the treatment of the infection due to H. pylori was statistically significant (p = 0.0297). The low percentage of H. pylori strains resistant to levofloxacin justify that the introduction of a triple therapy with levofloxacin is a good alternative in the treatment of H. pylori infections, especially in regions with high prevalence of H. pylori strains resistant to clarithromycin (> 20%)