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Development of a Highly Sensitive Method for Detection of Clarithromycin-Resistant Helicobacter pylori from Human Feces

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Abstract

Gastric infection of clarithromycin (CAM)-resistant Helicobacter pylori is one of the major causes of failure to eradicate this organism. A noninvasive and useful method for the detection of CAM-resistant H. pylori from human feces by restriction fragment length polymorphism (RFLP)-nested polymerase chain reaction (PCR) targeting the mutation of the 23S rRNA gene that confers CAM-resistance in H. pylori was developed in this study. Our nested PCR method detected DNA of H. pylori in feces with high sensitivity and specificity compared with both an enzyme-linked immunoadsorbent assay (ELISA) of H. pylori in feces and the isolation of H. pylori from gastric biopsy. Furthermore, the results of mutation analysis of the H. pylori 23S rRNA gene amplified from feces completely correlated with both that of the H. pylori 23S rRNA gene amplified from the isolates of gastric biopsy and the susceptibility of H. pylori isolates to CAM. Therefore, our results show that this RFLP/nested PCR method is useful for the accurate diagnosis of CAM-resistant H. pylori infection from feces.

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Literature Cited

  1. M Asaka K Satoh K Sugano T Sugiyama S Takahashi Y Fukuda et al. (2001) ArticleTitleGuidelines in the management of Helicobacter pylori infection in Japan Helicobacter 6 177–186 Occurrence Handle10.1046/j.1523-5378.2001.00027.x Occurrence Handle11683920

    Article  PubMed  Google Scholar 

  2. MJ Blaser (1996) ArticleTitleRole of vacA and the cagA locus of Helicobacter pylori in human disease Aliment Pharmacol Ther 10 IssueID(Suppl 1) 73–77

    Google Scholar 

  3. MP Dore MS Osato HM Malaty DY Graham (2000) ArticleTitleCharacterization of a culture method to recover Helicobacter pylori from the feces of infected patients Helicobacter 5 165–168 Occurrence Handle10.1046/j.1523-5378.2000.00026.x Occurrence Handle10971682

    Article  PubMed  Google Scholar 

  4. C Fontana M Favaro S Minelli AA Criscuolo A Pietroiusti A Galante et al. (2002) ArticleTitleNew site of modification of 23S rRNA associated with clarithromycin resistance of Helicobacter pylori clinical isolates Antimicrob Agents Chemother 46 3765–3769 Occurrence Handle10.1128/AAC.46.12.3765-3769.2002 Occurrence Handle12435674

    Article  PubMed  Google Scholar 

  5. C Fontana M Favaro A Pietroiusti ES Pistoia A Galante C Favalli (2003) ArticleTitleDetection of clarithromycin-resistant Helicobacter pylori in stool samples J Clin Microbiol 41 3636–3640 Occurrence Handle10.1128/JCM.41.8.3636-3640.2003 Occurrence Handle12904368

    Article  PubMed  Google Scholar 

  6. DY Graham GM Lew PD Klein DG Evans DJ Evans SuffixJr ZA Saeed SuffixJr et al. (1992) ArticleTitleEffect of treatment of Helicobacter pylori infection on the long-term recurrence of gastric or duodenal ulcer. A randomized, controlled study Ann Intern Med 116 705–708 Occurrence Handle1558340

    PubMed  Google Scholar 

  7. S Kolb C Knief S Stubner R Conrad (2003) ArticleTitleQuantitative detection of methanotrophs in soil by novel pmoA-targeted real-time PCR assays Appl Environ Microbiol 69 2423–2429 Occurrence Handle10.1128/AEM.69.5.2423-2429.2003 Occurrence Handle12732507

    Article  PubMed  Google Scholar 

  8. M Kurokawa M Minamide M Nukina H Nakanishi K Miki S Tomita et al. (1997) ArticleTitleUsefulness of Helicobacter pylori detection from feces specimens of the patients with peptic ulcer by polymerase chain reaction Kansenshogaku Zasshi 71 1168–1171 Occurrence Handle9455058

    PubMed  Google Scholar 

  9. JG Kusters MM Gerrits JA Strijp ParticleVan CM Vandenbroucke-Grauls (1997) ArticleTitleCoccoid forms of Helicobacter pylori are the morphologic manifestation of cell death Infect Immun 65 3672–3679 Occurrence Handle9284136

    PubMed  Google Scholar 

  10. DH Kwon MP Dore JJ Kim M Kato M Lee JY Wu et al. (2003) ArticleTitleHigh-level beta-lactam resistance associated with acquired multidrug resistance in Helicobacter pylori Antimicrob Agents Chemother 47 2169–2178 Occurrence Handle10.1128/AAC.47.7.2169-2178.2003 Occurrence Handle12821464

    Article  PubMed  Google Scholar 

  11. CT Loy LM Irwig PH Katelaris NJ Talley (1996) ArticleTitleDo commercial serological kits for Helicobacter pylori infection differ in accuracy? A meta-analysis Am J Gastroenterol 91 1138–1144 Occurrence Handle8651160

    PubMed  Google Scholar 

  12. A Makristathis E Pasching K Schutze M Wimmer ML Rotter AM Hirschl (1998) ArticleTitleDetection of Helicobacter pylori in stool specimens by PCR and antigen enzyme immunoassay J Clin Microbiol 36 2772–2774 Occurrence Handle9705436

    PubMed  Google Scholar 

  13. NP Mapstone DA Lynch FA Lewis AT Axon DS Tompkins MF Dixon et al. (1993) ArticleTitlePCR identification of Helicobacter pylori in faeces from gastritis patients Lancet 341 447 Occurrence Handle10.1016/0140-6736(93)93053-4

    Article  Google Scholar 

  14. H Miwa M Hirose S Kikuchi T Terai R Iwazaki O Kobayashi et al. (1999) ArticleTitleHow useful is the detection kit for antibody to Helicobacter pylori in urine (URINELISA) in clinical practice? Am J Gastroenterol 94 3460–3463 Occurrence Handle10.1111/j.1572-0241.1999.01608.x Occurrence Handle10606303

    Article  PubMed  Google Scholar 

  15. RA Moore B Beckthold S Wong A Kureishi LE Bryan (1995) ArticleTitleNucleotide sequence of the gyrA gene and characterization of ciprofloxacin-resistant mutants of Helicobacter pylori Antimicrob Agents Chemother 39 107–111 Occurrence Handle7695290

    PubMed  Google Scholar 

  16. National Committee for Clinical Laboratory Standards (2000) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, 5th edn. Approved Standard. Wayne, PA: National Committee for Clinical Laboratory Standards

  17. National Committee for Clinical Laboratory Standards (2002) Performance standards for antimicrobial susceptibility testing; twelfth informational supplement. M100-S12. Wayne, PA: National Committee for Clinical Laboratory Standards

  18. M Notarnicola F Russo A Cavallini M Bianco E Jirillo S Pece et al. (1996) ArticleTitlePCR identification of Helicobacter pylori DNA in faeces from patients with gastroduodenal pathology Med Sci Res 24 785–786

    Google Scholar 

  19. Y Okano KR Hristova CM Leutenegger LE Jackson RF Denison B Gebreyesus et al. (2004) ArticleTitleApplication of real-time PCR to study effects of ammonium on population size of ammonia-oxidizing bacteria in soil Appl Environ Microbiol 70 1008–1016 Occurrence Handle10.1128/AEM.70.2.1008-1016.2004 Occurrence Handle14766583

    Article  PubMed  Google Scholar 

  20. V Savarino S Vigneri G Celle (1999) ArticleTitleThe 13C urea breath test in the diagnosis of Helicobacter pylori infection Gut 45 IssueID(Suppl 1) 118–122

    Google Scholar 

  21. GG Stone D Shortridge RK Flamm J Versalovic J Beyer K Idler et al. (1996) ArticleTitleIdentification of a 23S rRNA gene mutation in clarithromycin-resistant Helicobacter pylori Helicobacter 1 227–228 Occurrence Handle9432314

    PubMed  Google Scholar 

  22. DE Taylor Z Ge D Purych T Lo K Hiratsuka (1997) ArticleTitleCloning and sequence analysis of two copies of a 23S rRNA gene from Helicobacter pylori and association of clarithromycin resistance with 23S rRNA mutations Antimicrob Agents Chemother 41 2621–2628 Occurrence Handle9420030

    PubMed  Google Scholar 

  23. JE Thomas GR Gibson MK Darboe A Dale LT Weaver (1992) ArticleTitleIsolation of Helicobacter pylori from human faeces Lancet 340 1194–1195 Occurrence Handle10.1016/0140-6736(92)92894-L Occurrence Handle1359263

    Article  PubMed  Google Scholar 

  24. CA Trieber DE Taylor (2002) ArticleTitleMutations in the 16S rRNA genes of Helicobacter pylori mediate resistance to tetracycline J Bacteriol 184 2131–2140 Occurrence Handle10.1128/JB.184.8.2131-2140.2002 Occurrence Handle11914344

    Article  PubMed  Google Scholar 

  25. N Uemura S Okamoto S Yamamoto N Matsumura S Yamaguchi M Yamakido et al. (2001) ArticleTitleHelicobacter pylori infection and the development of gastric cancer N Engl J Med 345 784–789 Occurrence Handle11556297

    PubMed  Google Scholar 

  26. D Vaira P Malfertheiner F Megraud AT Axon M Deltenre AM Hirschl et al. (1999) ArticleTitleDiagnosis of Helicobacter pylori infection with a new non-invasive antigen-based assay. HpSA European study group Lancet 354 30–33 Occurrence Handle10.1016/S0140-6736(98)08103-3 Occurrence Handle10406362

    Article  PubMed  Google Scholar 

  27. J Versalovic MS Osato K Spakovsky MP Dore R Reddy GG Stone et al. (1997) ArticleTitlePoint mutations in the 23S rRNA gene of Helicobacter pylori associated with different levels of clarithromycin resistance J Antimicrob Chemother 40 283–286 Occurrence Handle10.1093/jac/40.2.283 Occurrence Handle9301997

    Article  PubMed  Google Scholar 

  28. J Versalovic D Shortridge K Kibler MV Griffy J Beyer RK Flamm et al. (1996) ArticleTitleMutations in 23S rRNA are associated with clarithromycin resistance in Helicobacter pylori Antimicrob Agents Chemother 40 477–480 Occurrence Handle8834903

    PubMed  Google Scholar 

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Acknowledgments

We wish to thank Yuichi Jimbo, Shuji Soeda, and Hidenobu Kijima for their contributions to the study. This work was supported by a grant for private universities from the Ministry of Education, Science, Sports, and Culture, Japan, and by a grant from the Promotion and Mutual Aid Corporation for Private Schools of Japan.

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Correspondence to Norihisa Noguchi.

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Rimbara, E., Noguchi, N., Yamaguchi, T. et al. Development of a Highly Sensitive Method for Detection of Clarithromycin-Resistant Helicobacter pylori from Human Feces. Curr Microbiol 51, 1–5 (2005). https://doi.org/10.1007/s00284-004-4488-z

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  • DOI: https://doi.org/10.1007/s00284-004-4488-z

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