The occurrence of 16S rRNA gene mutations associated with resistance to tetracycline in H. pylori isolated in Bangladesh was investigated. Tetracycline susceptibility was determined by the agar dilution method. The 16S rRNA genes of these isolates were sequenced and analyzed. A tetracycline accumulation assay was performed. DNA sequence and transformation tests of nine tetracycline-resistant (MIC = 2 microg/ml) Bangladeshi H. pylori clinical isolates showed that in no case was the resistance due to mutations in the 16S rRNA gene, the only known cause of tetracycline resistance in this pathogen. Tetracycline accumulation assays implicated altered uptake or efflux.