Skip to main content
SpringerLink
Log in

Successful Culture Techniques for Helicobacter Species: General Culture Techniques for Helicobacter pylori

  • Protocol
  • First Online:
Helicobacter Species

Part of the book series: Methods in Molecular Biology ((MIMB,volume 921))

Abstract

Half of the world’s population is persistently infected with Helicobacter pylori. The chronicity of this infection ultimately elicits clinical manifestations ranging from gastritis and peptic ulcers to adenocarcinoma and MALT lymphoma. Laboratory research following the initial observations of Helicobacter species was greatly hindered by an inability to isolate and culture the bacteria. Thus, the ability to culture bacterial species from this genus is an extremely important step in expanding clinical knowledge and development of therapies. This chapter describes successful techniques for culturing H. pylori on selective horse blood agar media and in Brucella broth liquid media. Additionally, the specific growth requirements of other Helicobacter species are noted.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ernst PB, Gold BD (2000) The disease spectrum of Helicobacter pylori: the immunopathogenesis of gastroduodenal ulcer and gastric cancer. Annu Rev Microbiol 54:615–640

    Article  PubMed  CAS  Google Scholar 

  2. Baldwin DN, Shepherd B, Kraemer P et al (2007) Identification of Helicobacter pylori genes that contribute to stomach colonization. Infect Immun 75:1005–1016

    Article  PubMed  CAS  Google Scholar 

  3. Carpenter BM, McDaniel TK, Whitmire JM et al (2007) Expanding the Helicobacter pylori genetic toolbox: modification of an endogenous plasmid for use as a transcriptional reporter and complementation vector. Appl Environ Microbiol 73:7506–7514

    Article  PubMed  CAS  Google Scholar 

  4. Gancz H, Censini S, Merrell DS (2006) Iron and pH homeostasis intersect at the level of Fur regulation in the gastric pathogen Helicobacter pylori. Infect Immun 74: 602–614

    Article  PubMed  CAS  Google Scholar 

  5. Miles S, Piazuelo MB, Semino-Mora C et al (2010) Detailed in vivo analysis of the role of Helicobacter pylori Fur in colonization and disease. Infect Immun 78:3073–3082

    Article  PubMed  CAS  Google Scholar 

  6. Mouery K, Rader BA, Gaynor EC et al (2006) The stringent response is required for Helicobacter pylori survival of stationary phase, exposure to acid, and aerobic shock. J Bacteriol 188:5494–5500

    Article  PubMed  CAS  Google Scholar 

  7. Ottemann KM, Lowenthal AC (2002) Helicobacter pylori uses motility for initial colonization and to attain robust infection. Infect Immun 70:1984–1990

    Article  PubMed  CAS  Google Scholar 

  8. Salama N, Guillemin K, McDaniel TK et al (2000) A whole-genome microarray reveals genetic diversity among Helicobacter pylori strains. Proc Natl Acad Sci USA 97: 14668–14673

    Article  PubMed  CAS  Google Scholar 

  9. Fox JG, Dewhirst FE, Tully JG et al (1994) Helicobacter hepaticus sp. nov., a microaerophilic bacterium isolated from livers and intestinal mucosal scrapings from mice. J Clin Microbiol 32:1238–1245

    PubMed  CAS  Google Scholar 

  10. Fox JG, Rogers AB, Whary MT et al (2004) Helicobacter bilis-associated hepatitis in outbred mice. Comp Med 54:571–577

    PubMed  CAS  Google Scholar 

  11. Fox JG, Yan LL, Dewhirst FE et al (1995) Helicobacter bilis sp. nov., a novel Helicobacter species isolated from bile, livers, and intestines of aged, inbred mice. J Clin Microbiol 33:445–454

    PubMed  CAS  Google Scholar 

  12. Hodzic E, McKisic M, Feng S et al (2001) Evaluation of diagnostic methods for Helicobacter bilis infection in laboratory mice. Comp Med 51:406–412

    PubMed  CAS  Google Scholar 

  13. Mahler M, Bedigian HG, Burgett BL et al (1998) Comparison of four diagnostic ­methods for detection of Helicobacter species in laboratory mice. Lab Anim Sci 48:85–91

    PubMed  CAS  Google Scholar 

  14. Shames B, Fox JG, Dewhirst F et al (1995) Identification of widespread Helicobacter hepaticus infection in feces in commercial mouse colonies by culture and PCR assay. J Clin Microbiol 33:2968–2972

    PubMed  CAS  Google Scholar 

  15. Shomer NH, Dangler CA, Marini RP et al (1998) Helicobacter bilis/Helicobacter rodentium co-infection associated with diarrhea in a colony of scid mice. Lab Anim Sci 48:455–459

    PubMed  CAS  Google Scholar 

  16. Taylor NS, Xu S, Nambiar P et al (2007) Enterohepatic Helicobacter species are prevalent in mice from commercial and academic institutions in Asia, Europe, and North America. J Clin Microbiol 45:2166–2172

    Article  PubMed  CAS  Google Scholar 

  17. Blanchard TG, Nedrud JG (2006) Laboratory maintenance of Helicobacter species. In: Coico R, Kowalik T, Quarles JM et al (eds) Current protocols in microbiology. Wiley-Blackwell, Hoboken

    Google Scholar 

  18. Danon SJ, Lee A (2001) Other gastric helicobacters and spiral organisms. In: Mobley HLT, Mendz GL, Hazell SL (eds) Helicobacter pylori: physiology and genetics. ASM Press, Washington, DC

    Google Scholar 

  19. Maggio-Price L, Bielefeldt-Ohmann H, Treuting P et al (2005) Dual infection with Helicobacter bilis and Helicobacter hepaticus in p-glycoprotein-deficient mdr1a −/− mice results in colitis that progresses to dysplasia. Am J Pathol 166:1793–1806

    Article  PubMed  CAS  Google Scholar 

  20. Maggio-Price L, Shows D, Waggie K et al (2002) Helicobacter bilis infection accelerates and H. hepaticus infection delays the development of colitis in multiple drug resistance-deficient (mdr1a −/−) mice. Am J Pathol 160:739–751

    Article  PubMed  CAS  Google Scholar 

  21. Paster BJ, Lee A, Fox JG et al (1991) Phylogeny of Helicobacter felis sp. nov., Helicobacter mustelae, and related bacteria. Int J Syst Bacteriol 41:31–38

    Article  PubMed  CAS  Google Scholar 

  22. Ihrig M, Whary MT, Dangler CA et al (2005) Gastric Helicobacter infection induces a Th2 phenotype but does not elevate serum cholesterol in mice lacking inducible nitric oxide synthase. Infect Immun 73:1664–1670

    Article  PubMed  CAS  Google Scholar 

  23. Schauer DB (2001) Enterohepatic Helicobacter species. In: Mobley HLT, Mendz GL, Hazell SL (eds) Helicobacter pylori: physiology and genetics. ASM Press, Washington, DC

    Google Scholar 

  24. Dhaenens L, Szczebara F, Van Nieuwenhuyse S et al (1999) Comparison of iron uptake in different Helicobacter species. Res Microbiol 150:475–481

    Article  PubMed  CAS  Google Scholar 

  25. Douraghi M, Kashani SS, Zeraati H et al (2010) Comparative evaluation of three supplements for Helicobacter pylori growth in liquid culture. Curr Microbiol 60:254–262

    Article  PubMed  CAS  Google Scholar 

  26. Testerman TL, Conn PB, Mobley HL et al (2006) Nutritional requirements and antibiotic resistance patterns of Helicobacter species in chemically defined media. J Clin Microbiol 44:1650–1658

    Article  PubMed  CAS  Google Scholar 

  27. Senkovich O, Ceaser S, McGee DJ et al (2010) Unique host iron utilization mechanisms of Helicobacter pylori revealed with iron-deficient chemically defined media. Infect Immun 78:1841–1849

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Dr. Beth Carpenter for critical reading of the manuscript. Research in the laboratory of D. Scott Merrell is supported by R01AI065529 from NIH, R073PW from USUHS, and 300411-7.20-60393 from USMCI. The contents of this work are solely the responsibility of the authors and do not necessarily represent the official views of the NIH or Department of Defense (DoD).

Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD, USA

    Jeannette M. Whitmire & D. Scott Merrell

Authors
  1. Jeannette M. Whitmire
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. Scott Merrell
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D. Scott Merrell .

Editor information

Editors and Affiliations

  1. Medical School, Dept. Medicine, University of Massachusetts, Plantation Street 364, Worcester, 01635, Massachusetts, USA

    JeanMarie Houghton

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Whitmire, J.M., Merrell, D.S. (2012). Successful Culture Techniques for Helicobacter Species: General Culture Techniques for Helicobacter pylori . In: Houghton, J. (eds) Helicobacter Species. Methods in Molecular Biology, vol 921. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-005-2_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-005-2_4

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-004-5

  • Online ISBN: 978-1-62703-005-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation