Optimization of real-time PCR method to assess the direct sensitivity of clarithromycin in Helicobacter pylori
Subject Areas : Molecular Microbiology
Sadegh Ghorbani-Dalini
1
(Department of Microbiology, Jahrom Branch, Young Researcher’s Club, Islamic Azad University, Jahrom, Iran)
Mohammad Kargar
2
(Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran)
Abbas Doosti
3
(Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran)
Pejman Abbasi
4
(Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran)
Negar Souod
5
(Department of Microbiology, Jahrom Branch, Young Researcher’s Club, Islamic Azad University, Jahrom, Iran)
Keywords: Helicobacter pylori, Real-time PCR, Clarithromycin,
Abstract :
Background and Objectives: The increasing rate of clarithromycin resistance in H. pylori is recognized as a significant contributing factor for treatment failure. The aim of this study was to representing a real-time PCR assay in comparison with disk diffusion method for detection of clarithromycin resistance in H. pylori. Materials and Methods: This Experimental study was performed on 45 H. pylori specimens isolated from gastrointestinal disorders patients. In all samples, presence of H. pylori was confirmed by using culture, rapid urease test and conventional PCR. Also, clarithromycin resistance were assessed by using CLSI (Clinical and Laboratory Standard Institute) protocol and real-time PCR using specific primers and probe for H. pylori peptidylteransferas region of 23S rRNA gene. Results: Disk diffusion method identified 20 (44.44%) samples as susceptible and 25 (55.56%) samples as resistance to clarithromycin. But, real time PCR were identified samples as 20 (44.44%) with susceptible genotype, 5 (11.11%) with mix genotype and 20 (44.44%) as resistance to clarithromycin genotype. Also results showed that sensitivity of this assay was equal to 40 bacteria or 80 copy number of 23S rRNA gene. Conclusion: Results showed that Real-time PCR assay has high accuracy for identifying clarithromycin resistance in short time.
Fast and Accurate Quantitative Detection of Helicobacter pylori and Identification of Clarithromycin Resistance Mutations in H. pylori Isolates from Gastric Biopsy Specimens by Real-Time PCR,Prevalence of A2143G mutation of H. pylori-23S rRNAin Chinese subjects with and without clarithromycin use history,Evaluation of metronidazole resistance Helicobacterpylori strains isolated from Shahrekord,Evaluation of the Novel Helicobacter pylori ClariRes Real-Time PCR Assay for Detection and Clarithromycin Susceptibility Testing of H. pylori in Stool Specimens from Symptomatic Children,Real-Time PCR Assay for Rapid and Accurate Detection of Point Mutations Conferring Resistance to Clarithromycin in Helicobacter pylori,Real-Time PCR Assay Using Allele- Specific TaqMan probe for Detection of Clarithromycin Resistance and Its Point Mutations in Helicobacter Pylori,Role of Helicobacter pylori infection and nonsteroidal anti-inflammatory drugs in peptic-ulcer disease: a meta-analysis,The disease spectrum of Helicobacter pylori: the immunopathogenesis of gastroduodenal ulcer and gastric cancer,PCR-Based Diagnosis of Helicobacter pylori Infection and Real-Time Determination of Clarithromycin Resistance Directly from Human Gastric Biopsy Samples,Novel Real-Time PCR Assay for Detection of Helicobacter pylori Infection and Simultaneous Clarithromycin Susceptibility Testing of Stool and Biopsy Specimens,Rapid Detection of Mutations in the 23S rRNA Gene of Helicobacter pylori That Confers Resistance to Clarithromycin Treatment to the Bacterium,Direct Detection and Identification of Clarithromycin Resistance Point Mutations in Helicobacter pylori Strains Isolated from Biopsy Specimens by Real-Time PCR Assay,Detection of Clarithromycin-Resistant Helicobacter pylori in Stool Samples,Prevalence of A2143G, A2142G and A2142C mutations in producing Helicobacter pylori strains resistance to clarithromycin in Charmahal and Bakhtiari Province