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Liquid culture medium for the rapid cultivation of Helicobacter pylori from biopsy specimens

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Abstract

The goal of this study was to develop a liquid culture medium for the rapid isolation, cultivation, identification and subsequent antibiotics susceptibility testing of Helicobacter pylori directly from biopsy specimens. Five liquid media were tested: Ham’s F-12, Brucella broth, tryptic soybroth, brain heart infusion broth and Mueller-Hinton broth. After optimisation of the medium, it was applied in order to investigate biopsy samples from 150 patients with gastro-duodenal disorders and compared with traditional culture methods, microscopy and an H. pylori-specific TaqMan real-time polymerase chain reaction (PCR). The most reliable and rapid growth of H. pylori, even at a small inoculum size, was obtained in Ham’s F-12 medium with 5% horse serum. The developed system allowed the primary isolation of H. pylori in clinical samples and provided 87% sensitivity and 100% specificity.

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References

  1. NIH Consensus Conference (1994) Helicobacter pylori in peptic ulcer disease. JAMA 272:65–69

    Article  Google Scholar 

  2. Lee A (1995) Helicobacter infections in laboratory animals: a model for gastric neoplasias. Ann Med 27:575–582. doi:10.3109/07853899509002472

    Article  PubMed  CAS  Google Scholar 

  3. Talley NJ, Hunt RH (1997) What role does Helicobacter pylori play in dyspepsia and nonulcer dyspepsia? Arguments for and against H. pylori being associated with dyspeptic symptoms. Gastroenterology 113(Suppl 6):S67–S77

    PubMed  CAS  Google Scholar 

  4. Gilman RH (1990) Helicobacter pylori and gastritis in Peruvian patients: relationship to socioeconomic level, age, and sex. Am J Gastroenterol 85:819–823

    Google Scholar 

  5. Hegarty JP, Dowd MT, Baker KT (1999) Occurrence of Helicobacter pylori in surface water in the United States. J Appl Microbiol 87:697–701. doi:10.1046/j.1365-2672.1999.00912.x

    Article  PubMed  CAS  Google Scholar 

  6. Yingzhi L, Redlinger TE, Avitia R, Galindo A, Goodman K (2002) Isolation and genotyping of Helicobacter pylori from untreated municipal wastewater. Appl Environ Microbiol 68:1436–1439. doi:10.1128/AEM.68.3.1436-1439.2002

    Article  CAS  Google Scholar 

  7. Fendrick AM, Chernew ME, Hirth RA, Bloom BS, Bandekar RR, Scheiman JM (1999) Clinical and economic effects of population-based Helicobacter pylori screening to prevent gastric cancer. Arch Intern Med 159(2):142–148. doi:10.1001/archinte.159.2.142

    Article  PubMed  CAS  Google Scholar 

  8. Aguemon B, Struelens M, Deviere J, Denis O, Golstein P, Salmon I, Nagy N (2005) Primary antibiotic resistance and effectiveness of Helicobacter pylori triple therapy in ulcero-inflammatory pathologies of the upper digestive tract. Acta Gastroenterol Belg 68(3):287–293

    PubMed  Google Scholar 

  9. Buzás GM, Lotz G, Kiss A (2007) The Hungarian epidemiology of clarithromycin resistance in Helicobacter pylori infection. Orv Hetil 148(31):1461–1467. doi:10.1556/OH.2007.28013

    Article  PubMed  Google Scholar 

  10. Fischbach L, Evans EL (2007) Meta-analysis: the effect of antibiotic resistance status on the efficacy of triple and quadruple first-line therapies for Helicobacter pylori. Aliment Pharmacol Ther 26(3):343–357

    Article  PubMed  CAS  Google Scholar 

  11. Vakil N, Megraud F (2007) Eradication therapy for Helicobacter pylori. Gastroenterology 133(3):985–1001. doi:10.1053/j.gastro.2007.07.008

    Article  PubMed  CAS  Google Scholar 

  12. Han SW, Flamm R, Hachem C, Kim HY, Clarridge JE, Evans DG, Beyer J, Drnec J, Graham DY (1995) Transport and storage of Helicobacter pylori from gastric mucosal biopsies and clinical isolates. Eur J Clin Microbiol Infect Dis 14:349–352. doi:10.1007/BF02116531

    Article  PubMed  CAS  Google Scholar 

  13. Siu LK, Leung WK, Cheng AF, Sung JY, Ling TK, Ling JM, Ng EK, Lau JY, Chung SC (1998) Evaluation of a selective transport medium for gastric biopsy specimens to be cultured for Helicobacter pylori. J Clin Microbiol 36(10):3048–3050

    PubMed  CAS  Google Scholar 

  14. Soltesz V, Zeeberg B, Wadström T (1992) Optimal survival of Helicobacter pylori under various transport conditions. J Clin Microbiol 30:1453–1456

    PubMed  CAS  Google Scholar 

  15. Shahamat M, Mai U, Paszko-Kolva C, Yamamoto H, Colwell RR (1991) Evaluation of liquid media for growth of Helicobacter pylori. J Clin Microbiol 29:2835–2837

    PubMed  CAS  Google Scholar 

  16. Jiang X, Doyle MP (2000) Growth supplements for Helicobacter pylori. J Clin Microbiol 38(5):1984–1987

    PubMed  CAS  Google Scholar 

  17. Deshpande M, Calenoff E, Daniels L (1995) Rapid large-scale growth of Helicobacter pylori in flasks and fermentors. Appl Environ Microbiol 61(6):2431–2435

    PubMed  CAS  Google Scholar 

  18. Walsh EJ, Moran AP (1997) Influence of medium composition on the growth and antigen expression of Helicobacter pylori. J Appl Microbiol 83(1):67–75. doi:10.1046/j.1365-2672.1997.00164.x

    Article  PubMed  CAS  Google Scholar 

  19. Stevenson TH, Castillo A, Lucia LM, Acuff GR (2000) Growth of Helicobacter pylori in various liquid and plating media. Lett Appl Microbiol 30(3):192–196. doi:10.1046/j.1472-765x.2000.00699.x

    Article  PubMed  CAS  Google Scholar 

  20. Nedenskov P (1994) Nutritional requirements for growth of Helicobacter pylori. Appl Environ Microbiol 60(9):3450–3453

    PubMed  CAS  Google Scholar 

  21. Reynolds DJ, Penn CW (1994) Characteristics of Helicobacter pylori growth in a defined medium and determination of its amino acid requirements. Microbiology 140:2649–2656

    PubMed  CAS  Google Scholar 

  22. Albertson N, Wenngren I, Sjöström JE (1998) Growth and survival of Helicobacter pylori in defined medium and susceptibility to Brij 78. J Clin Microbiol 36(5):1232–1235

    PubMed  CAS  Google Scholar 

  23. Morshed MG, Karita M, Konishi H, Okita K, Nakazawa T (1994) Growth medium containing cyclodextrin and low concentration of horse serum for cultivation of Helicobacter pylori. Microbiol Immunol 38(11):897–900

    PubMed  CAS  Google Scholar 

  24. Kitsos CM, Stadtländer CT (1998) Helicobacter pylori in liquid culture: evaluation of growth rates and ultrastructure. Curr Microbiol 37(2):88–93. doi:10.1007/s002849900344

    Article  PubMed  CAS  Google Scholar 

  25. Vega AE, Cortiñas TI, Mattana CM, Silva HJ, Puig De Centorbi O (2003) Growth of Helicobacter pylori in medium supplemented with cyanobacterial extract. J Clin Microbiol 41(12):5384–5388. doi:10.1128/JCM.41.12.5384-5388.2003

    Article  PubMed  Google Scholar 

  26. Azevedo NF, Pacheco AP, Keevil CW, Vieira MJ (2004) Nutrient shock and incubation atmosphere influence recovery of culturable Helicobacter pylori from water. Appl Environ Microbiol 70(1):490–493. doi:10.1128/AEM.70.1.490-493.2004

    Article  PubMed  CAS  Google Scholar 

  27. Bury-Moné S, Kaakoush NO, Asencio C, Mégraud F, Thibonnier M, De Reuse H, Mendz GL (2006) Is Helicobacter pylori a true microaerophile? Helicobacter 11(4):296–303. doi:10.1111/j.1523-5378.2006.00413.x

    Article  PubMed  Google Scholar 

  28. Testerman TL, McGee DJ, Mobley H (2001) Helicobacter pylori growth and urease detection in the chemically defined medium Ham’s F-12 nutrient mixture. J Clin Microbiol 39(11):3842–3850. doi:10.1128/JCM.39.11.3842-3850.2001

    Article  PubMed  CAS  Google Scholar 

  29. Testerman TL, Conn PB, Mobley HL, McGee DJ (2006) Nutritional requirements and antibiotic resistance patterns of Helicobacter species in chemically defined media. J Clin Microbiol 44(5):1650–1658. doi:10.1128/JCM.44.5.1650-1658.2006

    Article  PubMed  CAS  Google Scholar 

  30. Greig MA, Neithercut WD, Hossack M, McColl KE (1991) Harnessing of urease activity of Helicobacter pylori to induce self-destruction of the bacterium. J Clin Pathol 44:157–159. doi:10.1136/jcp.44.2.157

    Article  PubMed  CAS  Google Scholar 

  31. Clyne M, Labigne A, Drumm B (1995) Helicobacter pylori requires an acidic environment to survive in the presence of urea. Infect Immun 63(5):1669–1673

    PubMed  CAS  Google Scholar 

  32. Sjöström JE, Larsson H (1996) Factors affecting growth and antibiotic susceptibility of Helicobacter pylori: effect of pH and urea on the survival of a wild-type strain and a urease-deficient mutant. J Med Microbiol 44(6):425–433

    Article  PubMed  Google Scholar 

  33. Xia HX, English L, Keane CT, O’Morain CA (1993) Enhanced cultivation of Helicobacter pylori in liquid media. J Clin Pathol 46:750–753. doi:10.1136/jcp.46.8.750

    Article  PubMed  CAS  Google Scholar 

  34. Morgan DR, Freedman R, Depew CE, Kraft WG (1987) Growth of Campylobacter pylori in liquid media. J Clin Microbiol 25:2123–2125

    PubMed  CAS  Google Scholar 

  35. Henriksen TH, Lia A, Schøyen R, Thoresen T, Berstad A (2000) Assessment of optimal atmospheric conditions for growth of Helicobacter pylori. Eur J Clin Microbiol Infect Dis 19(9):718–720. doi:10.1007/s100960000340

    Article  PubMed  CAS  Google Scholar 

  36. Kobayashi D, Eishi Y, Ohkusa T, Ishige I, Suzuki T, Minami J, Yamada T, Takizawa T, Koike M (2002) Gastric mucosal density of Helicobacter pylori estimated by real-time PCR compared with results of urea breath test and histological grading. J Med Microbiol 51:305–311

    PubMed  CAS  Google Scholar 

  37. Cattori V, Hofmann-Lehmann R (2008) Absolute quantitation of feline leukemia virus (FeLV) proviral DNA and viral RNA loads by TaqMan real-time PCR and RT-PCR. Methods Mol Biol 429:73–87

    Article  PubMed  CAS  Google Scholar 

  38. King A (2001) Recommendations for susceptibility tests on fastidious organisms and those requiring special handling. J Antimicrob Chemother 48(Suppl 1):77–80

    PubMed  CAS  Google Scholar 

  39. Ellerbrok H, Nattermann H, Ozel M, Beutin L, Appel B, Pauli G (2002) Rapid and sensitive identification of pathogenic and apathogenic Bacillus anthracis by real-time PCR. FEMS Microbiol Lett 214(1):51–59

    Article  PubMed  CAS  Google Scholar 

  40. Boddicker JD, Rota PA, Kreman T, Wangeman A, Lowe L, Hummel KB, Thompson R, Bellini WJ, Pentella M, Desjardin LE (2007) Real-time reverse transcription-PCR assay for detection of mumps virus RNA in clinical specimens. J Clin Microbiol 45(9):2902–2908. doi:10.1128/JCM.00614-07

    Article  PubMed  CAS  Google Scholar 

  41. Lindh M, Görander S, Andersson E, Horal P, Mattsby-Balzer I, Ryd W (2007) Real-time Taqman PCR targeting 14 human papilloma virus types. J Clin Virol 40(4):321–324. doi:10.1016/j.jcv.2007.09.009

    Article  PubMed  CAS  Google Scholar 

  42. Deltenre M, Glupczynski Y, De Prez C, Nyst JF, Burette A, Labbé M, Jonas C, DeKoster E (1989) The reliability of urease tests, histology and culture in the diagnosis of Campylobacter pylori infection. Scand J Gastroenterol Suppl 160:19–24. doi:10.3109/00365528909091730

    Article  PubMed  CAS  Google Scholar 

  43. Shadowen RD, Sciortino CV (1989) Improved growth of Campylobacter pylori in a biphasic system. J Clin Microbiol 27(8):1744–1747

    PubMed  CAS  Google Scholar 

  44. Frenck RW Jr, Clemens J (2003) Helicobacter in the developing world. Microbes Infect 5(8):705–713. doi:10.1016/S1286-4579(03)00112-6

    Article  PubMed  Google Scholar 

  45. National Committee for Clinical Laboratory Standards (NCCLS) (1999) Performance standards for antimicrobial susceptibility testing: 9th informational supplement: document M100-S9. NCCLS, Villanova, PA

  46. McNulty C, Owen R, Tompkins D, Hawtin P, McColl K, Price A, Smith G, Teare L; PHLS Helicobacter Working Group (2002) Helicobacter pylori susceptibility testing by disc diffusion. J Antimicrob Chemother 49(4):601–609. doi:10.1093/jac/49.4.601

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This study was performed with the financial support of the Swiss National Scientific Foundation (SNF) in the frameworks of the SCOPES 2005–2008 programme (IB73AO-110988/1).

We acknowledge the valuable contributions of Ion Cararus and Ion Chiriac for performing the endoscopic procedures.

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Authors and Affiliations

  1. Department of Microbiology, State University of Medicine and Pharmacy “N. Testemiţanu”, 165, bd. Stefan cel Mare si Sfint, 2004, Chishinau, Moldova

    N. Sainsus

  2. Clinical Laboratory, Vetsuisse Faculty, University of Zurich, Winterthurerstr. 260, 8057, Zurich, Switzerland

    V. Cattori & R. Hofmann-Lehmann

  3. Department of Surgery, Faculty of CME, Clinical Republican Hospital, State University of Medicine and Pharmacy “N. Testemiţanu”, Testemiţanu 29, 2025, Chishinau, Moldova

    C. Lepadatu

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  1. N. Sainsus
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  2. V. Cattori
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  3. C. Lepadatu
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  4. R. Hofmann-Lehmann
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Correspondence to N. Sainsus.

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Sainsus, N., Cattori, V., Lepadatu, C. et al. Liquid culture medium for the rapid cultivation of Helicobacter pylori from biopsy specimens. Eur J Clin Microbiol Infect Dis 27, 1209–1217 (2008). https://doi.org/10.1007/s10096-008-0567-6

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  • DOI: https://doi.org/10.1007/s10096-008-0567-6

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