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Comparison of three diagnostic assays for the identification of Helicobacter spp. in laboratory dogs

Abstract

A number of Helicobacter species may confound experimental data because of their association with disease progressing in various kinds of laboratory animals. Screening of Helicobacter species is particularly desirable, because they are prevalent in commercial and research animal facilities. The aim of the present study was to compare three diagnostic methods [e.g. Helicobacter stool antigen kit (HpSA), polymerase chain reaction (PCR) and rapid urease test (RUT)] for the identification of Helicobacter spp. in stools or gastric biopsy specimens collected from eight dogs suffering from gastritis. The gastroscopic biopsy specimens were tested using RUT and PCR, while stool specimens were evaluated using both HpSA and PCR. DNAs from the gastric biopsies and stool specimens were analyzed by both a consensus PCR that amplified the RNA polymerase beta-subunit-coding gene (rpoB) of Helicobacter spp. and a species-specific PCR to amplify the urease B gene of Helicobacter heilmannii, Helicobacter pylori, and Helicobacter felis. Helicobacter spp. were detected in 62.5% of the dogs, while H. heilmannii and H. felis were identified in 37.5 and 25% of the dogs, respectively. The HpSA did not efficiently detect Helicobacter spp. in the stool samples compared to the RUT and PCR assays, both of which successfully detected Helicobacter spp. in the two sample types. Finally, we recommend that consensus PCR with stool specimens could be used before the species-specific PCR for identifying Helicobacter species in laboratory dogs.

References

  1. 1.

    Vandamme P, Falsen E, Pot B, Kersters K, De Ley J. Identification of Campylobacter cinaedi isolated from blood and feces of children and adult females. J Clin Microbiol 1990; 28(5): 1016–1020.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  2. 2.

    Fox JG. The non-H pylori helicobacters: their expanding role in gastrointestinal and systemic diseases. Gut 2002; 50(2): 273–283.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  3. 3.

    Kim O. Helicobacter-An emerging new zoonotic pathogen. In: Zoonosis (Lorenzo-Morales J, ed), 1st ed, InTec, Rijeka, Croatia, 2012; pp 89–100.

    Google Scholar 

  4. 4.

    Ward JM, Fox JG, Anver MR, Haines DC, George CV, Collins MJ, Gorelick PL, Nagashima K, Gonda MA, Gilden RV, Tully JG, Russell RJ, Benveniste RE, Paster BJ, Dewhirst FE, Donovan JC, Anderson LM, Rice JM. Chronic active hepatitis and associated liver tumors in mice caused by a persistent bacterial infection with a novel Helicobacter species. J Natl Cancer Inst 1994; 86(16): 1222–1227.

    CAS  PubMed  Article  Google Scholar 

  5. 5.

    Eaton KA, Dewhirst FE, Paster BJ, Tzellas N, Coleman BE, Paola J, Sherding R. Prevalence and varieties of Helicobacter species in dogs from random sources and pet dogs: animal and public health implications. J Clin Microbiol 1996; 34(12): 3165–3170.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  6. 6.

    Chin EY, Dangler CA, Fox JG, Schauer DB. Helicobacter hepaticus infection triggers inflammatory bowel disease in T cell receptor alpha beta mutant mice. Comp Med 2000; 50(6): 586–594.

    CAS  PubMed  Google Scholar 

  7. 7.

    Shames B, Fox JG, Dewhirst F, Yan L, Shen Z, Taylor NS. Identification of widespread Helicobacter hepaticus infection in feces in commercial mouse colonies by culture and PCR assay. J Clin Microbiol 1995; 33(11): 2968–2972.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  8. 8.

    Goto K, Ohashi H, Takakura A, Itoh T. Current status of Helicobacter contamination of laboratory mice, rats, gerbils, and house musk shrews in Japan. Curr Microbiol 2000; 41(3): 161–166.

    CAS  PubMed  Article  Google Scholar 

  9. 9.

    Whary MT, Cline JH, King AE, Hewes KM, Chojnacky D, Salvarrey A, Fox JG. Monitoring sentinel mice for Helicobacter hepaticus, H rodentium, and H bilis infection by use of polymerase chain reaction analysis and serologic testing. Comp Med 2000; 50(4): 436–443.

    CAS  PubMed  Google Scholar 

  10. 10.

    Andersen LP, Norgaard A, Holck S, Blom J, Elsborg L. Isolation of a “Helicobacter heilmanii”-like organism from the human stomach. Eur J Clin Microbiol Infect Dis 1996; 15(1): 95–96.

    CAS  PubMed  Article  Google Scholar 

  11. 11.

    Solnick JV, O’Rourke J, Lee A, Paster BJ, Dewhirst FE, Tompkins LS. An uncultured gastric spiral organism is a newly identified Helicobacter in humans. J Infect Dis 1993; 168(2): 379–385.

    CAS  PubMed  Article  Google Scholar 

  12. 12.

    Solnick JV, O’Rourke J, Lee A, Tompkins LS. Molecular analysis of urease genes from a newly identified uncultured species of Helicobacter. Infect Immun 1994; 62(5): 1631–1638.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  13. 13.

    Mégraud F. The most important diagnostic modalities for Helicobacter pylori, now and in the future. Eur J Gastroenterol Hepatol 2012; 9(S1): S13–15.

    PubMed  Article  Google Scholar 

  14. 14.

    Leal YA, Flores LL, Fuentes-Pananá EM, Cedillo-Rivera R, Torres J. 13C-urea breath test for the diagnosis of Helicobacter pylori infection in children: a systematic review and meta-analysis. Helicobacter 2011; 16(4): 327–337.

    PubMed  Article  Google Scholar 

  15. 15.

    Nyan DC, Welch AR, Dubois A, Coleman WG Jr. Development of a noninvasive method for detecting and monitoring the time course of Helicobacter pylori infection. Infect Immun 2004; 72(9): 5358–5364.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  16. 16.

    Santos AM, Lopes T, Oleastro M, Chaves P, Cordeiro R, Ferreira M, Pereira T, Machado J, Guerreiro AS. Role of 13C-urea breath test in experimental model of Helicobacter pylori infection in mice. Helicobacter 2011; 16(4): 320–326.

    PubMed  Article  Google Scholar 

  17. 17.

    Neiger R, Dieterich C, Burnens A, Waldvogel A, Corthésy-Theulaz I, Halter F, Lauterburg B, Schmassmann A. Detection and prevalence of Helicobacter infection in pet cats. J Clin Microbiol 1998; 36(3): 634–637.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  18. 18.

    Kim S, Cho S, Kim O. Detection and identification of secreting Helicobacter species from cats. Lab Anim Res 2006; 22(3): 243–247.

    Google Scholar 

  19. 19.

    Monteiro L, Bonnemaison D, Vekris A, Petry KG, Bonnet J, Vidal R, Cabrita J, Mégraud F. Complex polysaccharides as PCR inhibitors in feces: Helicobacter pylori model. J Clin Microbiol 1997; 35(4): 995–998.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  20. 20.

    Moon DI, Shin EH, Oh HG, Oh JS, Hong S, Chung Y, Kim O. Usefulness of a Helicobacter pylori stool antigen test for diagnosing H. pylori infected C57BL/6 mice. Lab Anim Res 2013; 29(1): 27–32.

    PubMed  Article  Google Scholar 

  21. 21.

    Shimoyama T. Stool antigen tests for the management of Helicobacter pylori infection. World J Gastroenterol 2013; 19(45): 8188–8191.

    PubMed  PubMed Central  Article  Google Scholar 

  22. 22.

    Patel SK, Pratap CB, Jain AK, Gulati AK, Nath G. Diagnosis of Helicobacter pylori: what should be the gold standard? World J Gastroenterol 2014; 20(36): 12847–12859.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  23. 23.

    Lee H, Park Y, Kim O. Prevalence of Helicobacter Species in Feces of Dogs. Lab Anim Res 2007; 23(3): 339–344.

    Google Scholar 

  24. 24.

    Demirtürk L, Yazgan Y, Tarçin O, Ozel M, Diler M, Oncül O, Yildirim S. Does N-acetyl cystein affect the sensitivity and specificity of Helicobacter pylori stool antigen test? Helicobacter 2003; 8(2): 120–123.

    PubMed  Article  Google Scholar 

  25. 25.

    Hoshina S, Kahn SM, Jiang W, Green PH, Neu HC, Chin N, Morotomi M, LoGerfo P, Weinstein IB. Direct detection and amplification of Helicobacter pylori ribosomal 16S gene segments from gastric endoscopic biopsies. Diagn Microbiol Infect Dis 1990; 13(6): 473–479.

    CAS  PubMed  Article  Google Scholar 

  26. 26.

    Malfertheiner P1, Megraud F, O’Morain CA, Atherton J, Axon AT, Bazzoli F, Gensini GF, Gisbert JP, Graham DY, Rokkas T, El-Omar EM, Kuipers EJ; European Helicobacter Study Group. Management of Helicobacter pylori infection—the Maastricht IV/Florence Consensus Report. Gut 2012; 61(5): 646–664.

    CAS  PubMed  Article  Google Scholar 

  27. 27.

    Rautelin H, Lehours P, Mégraud F. Diagnosis of Helicobacter pylori infection. Helicobacter 2003; 8(S1): 13–20.

    CAS  PubMed  Article  Google Scholar 

  28. 28.

    Shinozaki JK, Sellon RK, Cantor GH, Besser TE, Mealey KL, Vaden SL. Fecal polymerase chain reaction with 16S ribosomal RNA primers can detect the presence of gastrointestinal Helicobacter in dogs. J Vet Intern Med 2002; 16(4): 426–432.

    PubMed  Article  Google Scholar 

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Acknowledgments

This study was supported by a Wonkwang University research grant in 2015. We wish to thank Gi-Wook Oh, (Center for Animal Resources Development, Wonkwang University) for technical support.

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Correspondence to Okjin Kim.

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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Hong, S., Chung, Y., Kang, W. et al. Comparison of three diagnostic assays for the identification of Helicobacter spp. in laboratory dogs. Lab Anim Res 31, 86–92 (2015). https://doi.org/10.5625/lar.2015.31.2.86

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Keywords

  • Helicobacter
  • stool antigen kit
  • stool
  • PCR
  • dog