title: Laboratory handling of Helicobacter pylori critically influences the results of in-vitro metronidazole resistance determination.
abstract: In-vitro metronidazole resistance rates of Helicobacter pylori determined by Etest are high, and the predictive value of metronidazole resistance is low. It was hypothesised that altered laboratory methods could reduce the overestimation of resistance and improve the predictive value of the Etest. Pre-treatment isolates (n = 150) of H. pylori from 150 patients were investigated by Etest with incubation for 72 h. Treatment with metronidazole, tetracycline and bismuth for 10 days failed to eradicate H. pylori in 23 patients. After isolate storage for 3 years, resistance determination results by agar dilution and Etest, with incubation for 72 and 31 h, were compared. The rate of metronidazole resistance was reduced significantly during storage, and instability of resistance was associated significantly with treatment outcome. Isolates that retained in-vitro resistance had significantly (p 0.008) higher treatment failure rates (n = 13; 42%) than isolates that lost resistance (n = 3; 9%). The reproducibility achieved by dual testing with agar dilution and Etest was 41% and 70% for +/- 1 and +/- 2 log2 dilutions, respectively, after incubation for 72 h, and 85% and 92%, respectively, after incubation for 31 h. Thus, the predictive value was improved from 25% to 50% by the altered laboratory conditions (p 0.04). MIC values of 2-8 mg/L signified an intermediate risk of treatment failure.
authors.affiliation: Department of Microbiology, Sentralsykehuset i Vestfold, Tønsberg, Norway. thor-henrik.henriksen@helse-bergen.no
authors.forename: T H
authors.initials: TH
authors.lastname: Henriksen
authors.forename: F
authors.initials: F
authors.lastname: Lerang
authors.forename: A
authors.initials: A
authors.lastname: Lia
authors.forename: R
authors.initials: R
authors.lastname: Schøyen
authors.forename: T
authors.initials: T
authors.lastname: Thoresen
authors.forename: T
authors.initials: T
authors.lastname: Berge
authors.forename: E
authors.initials: E
authors.lastname: Ragnhildstveit
authors.forename: Y
authors.initials: Y
authors.lastname: Tveten
authors.forename: A
authors.initials: A
authors.lastname: Berstad
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000890:Anti-Infective Agents / Q000494:pharmacology* / Q000627:therapeutic use; D001729:Bismuth / Q000627:therapeutic use; D024881:Drug Resistance, Bacterial; D004359:Drug Therapy, Combination; D005260:Female; D016481:Helicobacter Infections / Q000188:drug therapy / Q000382:microbiology; D016480:Helicobacter pylori / Q000187:drug effects*; D006801:Humans; D008297:Male; D008795:Metronidazole / Q000494:pharmacology* / Q000627:therapeutic use; D008826:Microbial Sensitivity Tests / Q000379:methods; D008875:Middle Aged; D010118:Oxytetracycline / Q000627:therapeutic use; D013048:Specimen Handling / Q000379:methods*; D016896:Treatment Outcome
chemical_list: D000890:Anti-Infective Agents; D008795:Metronidazole; D001729:Bismuth; D010118:Oxytetracycline