Prevalence of Helicobacter pylori in symptomatic patients and detection of clarithromycin resistance using melting curve analysis

Ayse Demet Kaya; C Elif Oztürk; Yusuf Akcan; Mustafa Behçet; A Esra Karakoç; Mihriban Yücel; Müge Mısırlıoglu; Serdar Tuncer

doi:10.1016/j.curtheres.2007.06.001

Prevalence of Helicobacter pylori in symptomatic patients and detection of clarithromycin resistance using melting curve analysis

Curr Ther Res Clin Exp. 2007 May;68(3):151-60. doi: 10.1016/j.curtheres.2007.06.001.

Authors

Ayse Demet Kaya¹, C Elif Oztürk¹, Yusuf Akcan², Mustafa Behçet¹, A Esra Karakoç³, Mihriban Yücel³, Müge Mısırlıoglu⁴, Serdar Tuncer⁴

Affiliations

¹ Department of Microbiology, Duzce University, Medical Faculty Hospital, Konuralp/Duzce, Turkey.
² Department of Gastroenterology, Duzce University, Medical Faculty Hospital, Konuralp/Duzce, Turkey.
³ Department of Microbiology, Ministry of Health, Ankara Training and Research Hospital, Ankara, Turkey.
⁴ Metis Biotechnology Laboratory, Ankara, Turkey.

Abstract

Abstract.

Background: Clarithromycin is often a component of combination therapies for Helicobacter pylori eradication; however, increases in resistance rates have decreased the success of the treatment.

Objective: This study was designed to determine the prevalence of H pylori infection in symptomatic patients and to detect clarithromycin resistance rates using melting curve analysis.

Methods: Patients scheduled for upper endoscopy at the Endoscopy Unit of the Department of Gastroenterology, Duzce University, Medical Faculty Hospital, Konuralp/Duzce, Turkey, were assessed for enrollment in the study. Two pairs of gastric biopsy specimens (antrum and corpus) were obtained from each study patient. Histopathologic examination, rapid urease test, culture, and polymerase chain reaction (PCR) of the specimens were used to identify H pylori infection. Clarithromycin resistance was detected using melting curve analysis.

Results: Seventy-five patients (41 women, 34 men; mean [SD]age, 42.6 [14.5] years [range, 17-70 years]) were included in the study. Using histopathology and rapid urease test, H pylori was detected in 40 (53.3%) of the 75 specimens. H pylori was detected using PCR in 40 (53.3%) specimens and by culture in 10 (13.3%) specimens. The specificity and sensitivity of PCR and culture were interpreted by comparing them with the results of histopathologic examination and urease tests. The specificity and sensitivity of PCR were 68.6% and 72.5%, respectively, and the specificity and sensitivity of culture were 97.1% and 22.5%, respectively. Of the 40 isolates, 21 (52.5%) were susceptible to clarithromycin, 12 (30.0%) were resistant, and a mixed susceptibility pattern was detected in 7 (17.5%) specimens. H pylori isolates from 19 (79.2%) of the 24 patients who had formerly used clarithromycin showed clarithromycin resistance.

Conclusions: The prevalence of H pylori infection was 53.3% for the symptomatic patients in this study, and 47.5% of the isolates showed clarithromycin resistance using melting curve analysis. The PCR-based system used in this study was accurate for the detection of H pylori infection as well as clarithromycin susceptibility testing directly in biopsy specimens.

Keywords: Helicobacter pylori; PCR; clarithromycin resistance; melting curve analysis.