Bacterial species genotyping in the Helicobacter pylori refered patients to hospitals of Tehran using RFLP technique

Background and Aims: Helicobacter pylori is one of the most common human pathogens and since it was discovered in ۱۹۸۲ by Marshall and Warren has been established as one of the most variable and diverse bacteria known to man. Resident to the lining of the human stomach it is seen as the cause of a multitude of illnesses, ranging from mild gastritis, gastroduodenal ulcers as well as gastric cancer and controversially even heart disease.materials and methods: The differentiation of a pathogen so prevalent and variable presents a challenge. To date two versions of Helicobacter pylori have been sequenced indicating an immense variability in its genome. To differentiate the Helicobacter pylori subtypes, the PCR-RFLP technique was used (polymerase chain reaction used as amplification and restricted fragment length polymorphism used as differentiation method). We propose that from ۳۷ Helicobacter pylori positive gastric biopsies and ۳۳ viable PCR products the selection of two different genes (UreAB and UreCD) and PCR-RFLP on these two genes (using HaeIII and NdeII respectively) is superior to PCR-RFLP on only one gene. After running the experiment, we include an analysis of the resultant gel electrophoresis band patterns with visual evaluation.Results: Two combined PCR-RFLP runs was superior to both of the single runs with only one pair of Helicobacter pylori strains showing an identical band pattern in both UreAB/HaeIII and UreCD/NdeII runs.Conclusion: This would allow the clinician to make a more precise judgment on which patient it is necessary to treat, therefore reducing costs to society and slowing down the emergence of resistance to antibiotics.

Keywords:

genotyping , Helicobacter pylori , RFLP technique