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.
Key Words: Helicobacter pylori, clarithromycin resistance, melting curve analysis, PCR
Full Text
The Full Text of this article is available as a PDF (687.9 KB).
References
- 1.Lee MG, Arthurs M, Smikle MF. Antibiotic sensitivity of Helicobacter pylori in Jamaica. West Indian Med J. 2004;53:374–377. [PubMed] [Google Scholar]
- 2.Kaneko F, Suzuki H, Hasegawa N. High prevalence rate of Helicobacter pylori resistance to clarithromycin during long-term multiple antibiotic therapy for chronic respiratory disease caused by non-tuberculous mycobacteria. Aliment Pharmacol Ther. 2004;20(Suppl 1):62–67. doi: 10.1111/j.1365-2036.2004.01993.x. [DOI] [PubMed] [Google Scholar]
- 3.Glupczynski Y, Mégraud F, Lopez-Brea M, Andersen LP. European multicentre survey of in vitro antimicrobial resistance in Helicobacter pylori. Eur J Clin Microbiol Infect Dis. 2001;20:820–823. doi: 10.1007/s100960100611. [DOI] [PubMed] [Google Scholar]
- 4.McLoughlin R, Racz I, Buckley M. Therapy of Helicobacter pylori. Helicobacter. 2004;9(Suppl 1):42–48. doi: 10.1111/j.1083-4389.2004.00251.x. [DOI] [PubMed] [Google Scholar]
- 5.Soltermann A, Perren A, Schmid S. Assessment of Helicobacter pylori clarithromycin resistance mutations in archival gastric biopsy samples. Swiss Med Wkly. 2005;135:327–332. doi: 10.4414/smw.2005.10866. [DOI] [PubMed] [Google Scholar]
- 6.Oleastro M, Menard A, Santos A. Real-time PCR assay for rapid and accurate detection of point mutations conferring resistance to clarithromycin in Helicobacter pylori. J Clin Microbiol. 2003;41:397–402. doi: 10.1128/JCM.41.1.397-402.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7.Velapatino B, Balqui J, Gilman RH. Validation of string test for diagnosis of Helicobacter pylori infections. J Clin Microbiol. 2006;44:976–980. doi: 10.1128/JCM.44.3.976-980.2006. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8.Valentine JL. PCR detection of Helicobacter pylori. In: Persing DH, Smith TF, Tenover FC, White TJ, editors. Diagnostic Molecular Microbiology Principles and Applications. 1st ed. American Society for Microbiology; Washington, DC: 1993. pp. 284–287. [Google Scholar]
- 9.Chisholm SA, Owen RJ, Teare EL, Saverymuttu S. PCR-based diagnosis of Helicobacter pylori infection and real-time determination of clarithromycin resistance directly from human gastric biopsy samples. J Clin Microbiol. 2001;39:1217–1220. doi: 10.1128/JCM.39.4.1217-1220.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Yilmaz O, Sen N, Kupelioglu AA, Simsek I. Detection of H pylori infection by ELISA and Western blot techniques and evaluation of anti CagA seropositivity in adult Turkish dyspeptic patients. World J Gastroenterol. 2006;12:5375–5378. doi: 10.3748/wjg.v12.i33.5375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11.Dunn BE, Cohen H, Blaser MJ. Helicobacter pylori. Clin Microbiol Rev. 1997;10:720–741. doi: 10.1128/cmr.10.4.720. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12.Kisioglu AN, Birer S, Saribas S. Helicobacter pylori infeksiyonlarının serolojik tanısında Western Blot yönteminin degeri. Infeksiyon Derg. 2004;18:467–472. [Google Scholar]
- 13.Demiray E, Yilmaz O, Sarkis C. Comparison of invasive methods and two different stool antigen tests for diagnosis of H pylori infection in patients with gastric bleeding. World J Gastroenterol. 2006;12:4206–4210. doi: 10.3748/wjg.v12.i26.4206. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14.Blaser MJ. Helicobacter pylori and other gastric Helicobacter species. In: Mandell GL, Douglas RG, Bennett JE, editors. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases. 5th ed. Churchill Livingstone; Philadelphia, Pa: 2000. pp. 2285–2293. [Google Scholar]
- 15.Lo CC, Lai KH, Peng NJ. Polymerase chain reaction: A sensitive method for detecting Helicobacter pylori infection in bleeding peptic ulcers. World J Gastroenterol. 2005;11:3909–3914. doi: 10.3748/wjg.v11.i25.3909. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16.Baglan PH, Bozdayi G, Ozkan M. Clarithromycin resistance prevalence and Icea gene status in Helicobacter pylori clinical isolates in Turkish patients with duodenal ulcer and functional dyspepsia. J Microbiol. 2006;44:409–416. [PubMed] [Google Scholar]
- 17.Versalovic J, Shortridge D, Kibler K. Mutations in 23S rRNA are associated with clarithromycin resistance in Helicobacter pylori. Antimicrob Agents Chemother. 1996;40:477–480. doi: 10.1128/aac.40.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18.van Doorn LJ, Glupczynski Y, Kusters JG. Accurate prediction of macrolide resistance in Helicobacter pylori by a PCR line probe assay for detection of mutations in the 23S rRNA gene: Multicenter Validation Study. Antimicrob Agents Chemother. 2001;45:1500–1504. doi: 10.1128/AAC.45.5.1500-1504.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19.Lascols C, Lamarque D, Costa JM. 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. J Clin Microbiol. 2003;41:4573–4577. doi: 10.1128/JCM.41.10.4573-4577.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20.Posteraro P, Branca G, Sanguinetti M. Rapid detection of clarithromycin resistance in Helicobacter pylori using a PCR-based denaturing HPLC assay. J Antimicrob Chemother. 2006;57:71–78. doi: 10.1093/jac/dki406. [DOI] [PubMed] [Google Scholar]
- 21.Takahashi S, Tokunaga K, Kai A. The antimicrobial susceptibility test of Helicobacter pylori [in Japanese] Nippon Rinsho. 2003;61:79–83. [PubMed] [Google Scholar]
- 22.Goldman RC, Zakula D, Flamm R. Tight binding of clarithromycin, its 14-(R)-hydroxy metabolite, and erythromycin to Helicobacter pylori ribosomes. Antimicrob Agents Chemother. 1994;38:1496–1500. doi: 10.1128/aac.38.7.1496. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 23.Graham DY, Lew GM, Klein PD. Effect of treatment of Helicobacter pylori infection on the long-term recurrence of gastric or duodenal ulcer. A randomized, controlled study. Ann Intern Med. 1992;116:705–708. doi: 10.7326/0003-4819-116-9-705. [DOI] [PubMed] [Google Scholar]
- 24.Kim JM, Kim JS, Jung HC. Distribution of antibiotic MICs for Helicobacter pylori strains over a 16-year period in patients from Seoul, South Korea. Antimicrob Agents Chemother. 2004;48:4843–4847. doi: 10.1128/AAC.48.12.4843-4847.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 25.Megraud F. H pylori antibiotic resistance: Prevalence, importance, and advances in testing. Gut. 2004;53:1374–1384. doi: 10.1136/gut.2003.022111. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26.Cheng H, Hu FL. The epidemiology of Helicobacter pylori resistance to antibiotics in Beijing [in Chinese] Zhonghua Yi Xue Za Zhi. 2005;85:2754–2757. [PubMed] [Google Scholar]
- 27.Perez Aldana L, Kato M, Nakagawa S. The relationship between consumption of antimicrobial agents and the prevalence of primary Helicobacter pylori resistance. Helicobacter. 2002;7:306–309. doi: 10.1046/j.1523-5378.2002.00096.x. [DOI] [PubMed] [Google Scholar]
- 28.Kolaylı F, Karadenizli A, Çelebi A, Bingöl R. Helicobacter pylori suslarının metronidazol, klaritromisin ve amoksisiline in vitro duyarlılıkları. Infeksiyon Derg. 2004;18:473–476. [Google Scholar]
- 29.Simsek H, Balaban YH, Gunes DD. Alarming clarithromycin resistance of Helicobacter pylori in Turkish population. Helicobacter. 2005;10:360–361. doi: 10.1111/j.1523-5378.2005.00325.x. [DOI] [PubMed] [Google Scholar]
- 30.Can F, Demirbilek M, Selcuk H. Clarithromycin resistance of Helicobacter pylori strains isolated from antral biopsy specimens [in Turkish] Mikrobiyol Bul. 2004;38:349–353. [PubMed] [Google Scholar]