Author/Authors :
Dorafshan، Shima نويسنده Department of Microbiology, Faculty of Basic Sciences, Qom Branch of Islamic Azad University, Qom, IR Iran Dorafshan, Shima , Alebouyeh، Masoud نويسنده , , Shokrzadeh، Leila نويسنده , , Mirzaei، Tabasom نويسنده Department of Microbiology, Faculty of Basic Sciences, Qom Branch of Islamic Azad University, Qom, IR Iran Mirzaei, Tabasom , Nazemalhosseini Mojarad، Ehsan نويسنده Gastroenterology and liver Diseases Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran Nazemalhosseini Mojarad, Ehsan , Behzad، Catherine نويسنده Gastroenterology and liver Diseases Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran Behzad, Catherine , Miri، Marzyieh نويسنده Gastroenterology and liver Diseases Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran Miri, Marzyieh , Taghavi، Yaser نويسنده Gastroenterology and liver Diseases Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran Taghavi, Yaser , zojaji، homayoun نويسنده shahid beheshti university , , Asadzadeh Aghdaei، Hamid نويسنده Basic and Molecular epidemiology of Gastrointestinal Disorders Research Center, Shahid Beheshti University Medical Sciences, Tehran , , Zali، Mohamad Reza نويسنده Research Center for Gastroenterology and Liver Disease, Shahid Beheshti University of Medical Sciences, Tehran ,
Abstract :
Background: Rapid detection of resistant strains of Helicobacter pylori in human clinical samples is of major importance in clinical
settings. Inability of conventional clinical laboratory techniques in detection of these strains usually leads to failure of prescribed
therapeutic regimens. Objectives: The aim of this study was designing a simple and rapid allele-specific PCR (AS-PCR)-based method for detection of more
frequent gyrA mutations at Asn87Lys codon, responsible for emergence of fluoroquinolone resistance in H. pylori strains. Patients and Methods: All bacterial strains were obtained from clinical biopsy samples in our laboratory. Identification of the isolates was
performed by the genus- and species-specific primers and allele-specific primers, designed to match with the site of the point mutations.
Samples with positive results for the designed PCR method were sequenced to verify the existence of the target mutations. Results: Point mutations in the gyrA gene at Asn87Lys codon (AAT > AAA and AAC > AAG) were detected in all standard resistant strains as
well as some of clinical isolates with previously determined resistance phenotypes for fluoroquinolones. Presence of the target mutations
was successfully confirmed in all the control strains by the newly designed primers and sequencing. Conclusions: The designed AS-PCR was a good and reliable method for detection of AAT > AAA and AAC > AAG point mutations in H. pylori
isolates.
