Assessment of metronidazole susceptibility in Helicobacter pylori: statistical validation and error rate analysis of breakpoints determined by the disk diffusion test.
Metronidazole susceptibility of 100 Helicobacter pylori strains was assessed by determining the inhibition zone diameters by disk diffusion test and the MICs by agar dilution and PDM Epsilometer test (E test). Linear regression analysis was performed, allowing the definition of significant linear relations, and revealed correlations of disk diffusion results with both E-test and agar dilution results (r2 = 0.88 and 0.81, respectively). No significant differences (P = 0.84) were found between MICs defined by E test and those defined by agar dilution, taken as a standard. Reproducibility comparison between E-test and disk diffusion tests showed that they are equivalent and with good precision. Two interpretative susceptibility schemes (with or without an intermediate class) were compared by an interpretative error rate analysis method. The susceptibility classification scheme that included the intermediate category was retained, and breakpoints were assessed for diffusion assay with 5-microg metronidazole disks. Strains with inhibition zone diameters less than 16 mm were defined as resistant (MIC > 8 microg/ml), those with zone diameters equal to or greater than 16 mm but less than 21 mm were considered intermediate (4 microg/ml < MIC </= 8 microg/ml), and those with zone diameters of 21 mm or greater were regarded as susceptible (MIC </= 4 microg/ml). Error rate analysis applied to this classification scheme showed occurrence frequencies of 1% for major errors and 7% for minor errors, when the results were compared to those obtained by agar dilution. No very major errors were detected, suggesting that disk diffusion might be a good alternative for determining the metronidazole sensitivity of H. pylori strains.
Chaves
S
S
Laboratório de Bacteriologia, Instituto Nacional de Saúde, Lisbon, Portugal.
Gadanho
M
M
Tenreiro
R
R
Cabrita
J
J
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004058:Diffusion; D016480:Helicobacter pylori / Q000187:drug effects*; D008795:Metronidazole / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests / Q000379:methods*; D012044:Regression Analysis; D015203:Reproducibility of Results
D000900:Anti-Bacterial Agents; D008795:Metronidazole