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. 1996 Jan;3(1):73–78. doi: 10.1128/cdli.3.1.73-78.1996

A sensitive and specific PCR method to detect Helicobacter felis in a conventional mouse model.

L Kong 1, J G Smith 1, D Bramhill 1, G K Abruzzo 1, C Bonfiglio 1, C Cioffe 1, A M Flattery 1, C J Gill 1, L Lynch 1, P M Scott 1, L Silver 1, C Thompson 1, H Kropp 1, K Bartizal 1
PMCID: PMC170250  PMID: 8770507

Abstract

Although many detection methods have been used to determine Helicobacter colonization in small animal models, the sensitivity and specificity of these detection methods are limited. To improve the Helicobacter felis conventional mouse model for accurate evaluation of therapeutic regimens, we developed a PCR for detection of, and a competitive PCR for quantitation of, H. felis in viral antibody-free (VAF) mice. The PCR was based on the H. felis 16S rRNA gene. An internal control DNA was used for competitive quantitation of the PCR. VAF conventional Swiss-Webster mice were infected with an H. felis culture by oral gavage. At various times after H. felis challenge and therapy, stomach mucosa was collected and evaluated by PCR. PCR detected approximately 50 to 100 H. felis cells per mouse stomach and showed no cross-reaction with other bacteria commonly found in mouse stomachs. Colonization of H. felis in the mouse stomach was confirmed by culture isolation from germfree mice and histological examination of VAF mice. Response to therapy in this H. felis model correlated well with results seen in human clinical trials with H. pylori. A model utilizing PCR detection which may be useful for discovering new antibiotics and/or vaccines against Helicobacter ulcer disease has been developed.

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Selected References

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  1. Baskerville A., Newell D. G. Naturally occurring chronic gastritis and C pylori infection in the rhesus monkey: a potential model for gastritis in man. Gut. 1988 Apr;29(4):465–472. doi: 10.1136/gut.29.4.465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell G. D., Powell K. U. Eradication of Helicobacter pylori and its effect in peptic ulcer disease. Scand J Gastroenterol Suppl. 1993;196:7–11. doi: 10.3109/00365529309098334. [DOI] [PubMed] [Google Scholar]
  3. Clayton C., Kleanthous K., Tabaqchali S. Detection and identification of Helicobacter pylori by the polymerase chain reaction. J Clin Pathol. 1991 Jun;44(6):515–516. doi: 10.1136/jcp.44.6.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dick-Hegedus E., Lee A. Use of a mouse model to examine anti-Helicobacter pylori agents. Scand J Gastroenterol. 1991 Sep;26(9):909–915. doi: 10.3109/00365529108996241. [DOI] [PubMed] [Google Scholar]
  5. Eaton K. A., Morgan D. R., Krakowka S. Persistence of Helicobacter pylori in conventionalized piglets. J Infect Dis. 1990 Jun;161(6):1299–1301. doi: 10.1093/infdis/161.6.1299. [DOI] [PubMed] [Google Scholar]
  6. Eckloff B. W., Podzorski R. P., Kline B. C., Cockerill F. R., 3rd A comparison of 16S ribosomal DNA sequences from five isolates of Helicobacter pylori. Int J Syst Bacteriol. 1994 Apr;44(2):320–323. doi: 10.1099/00207713-44-2-320. [DOI] [PubMed] [Google Scholar]
  7. Fox J. G., Yan L. L., Dewhirst F. E., Paster B. J., Shames B., Murphy J. C., Hayward A., Belcher J. C., Mendes E. N. Helicobacter bilis sp. nov., a novel Helicobacter species isolated from bile, livers, and intestines of aged, inbred mice. J Clin Microbiol. 1995 Feb;33(2):445–454. doi: 10.1128/jcm.33.2.445-454.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goodwin C. S., Blincow E., Peterson G., Sanderson C., Cheng W., Marshall B., Warren J. R., McCulloch R. Enzyme-linked immunosorbent assay for Campylobacter pyloridis: correlation with presence of C. pyloridis in the gastric mucosa. J Infect Dis. 1987 Mar;155(3):488–494. doi: 10.1093/infdis/155.3.488. [DOI] [PubMed] [Google Scholar]
  9. Ho S. A., Hoyle J. A., Lewis F. A., Secker A. D., Cross D., Mapstone N. P., Dixon M. F., Wyatt J. I., Tompkins D. S., Taylor G. R. Direct polymerase chain reaction test for detection of Helicobacter pylori in humans and animals. J Clin Microbiol. 1991 Nov;29(11):2543–2549. doi: 10.1128/jcm.29.11.2543-2549.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kanangat S., Solomon A., Rouse B. T. Use of quantitative polymerase chain reaction to quantitate cytokine messenger RNA molecules. Mol Immunol. 1992 Oct;29(10):1229–1236. doi: 10.1016/0161-5890(92)90059-7. [DOI] [PubMed] [Google Scholar]
  11. Karita M., Li Q., Cantero D., Okita K. Establishment of a small animal model for human Helicobacter pylori infection using germ-free mouse. Am J Gastroenterol. 1994 Feb;89(2):208–213. [PubMed] [Google Scholar]
  12. Karita M., Li Q., Okita K. Evaluation of new therapy for eradication of H. pylori infection in nude mouse model. Am J Gastroenterol. 1993 Sep;88(9):1366–1372. [PubMed] [Google Scholar]
  13. Lee A., Fox J. G., Otto G., Murphy J. A small animal model of human Helicobacter pylori active chronic gastritis. Gastroenterology. 1990 Nov;99(5):1315–1323. doi: 10.1016/0016-5085(90)91156-z. [DOI] [PubMed] [Google Scholar]
  14. Lee A., Phillips M. W., O'Rourke J. L., Paster B. J., Dewhirst F. E., Fraser G. J., Fox J. G., Sly L. I., Romaniuk P. J., Trust T. J. Helicobacter muridarum sp. nov., a microaerophilic helical bacterium with a novel ultrastructure isolated from the intestinal mucosa of rodents. Int J Syst Bacteriol. 1992 Jan;42(1):27–36. doi: 10.1099/00207713-42-1-27. [DOI] [PubMed] [Google Scholar]
  15. Malfertheiner P. Compliance, adverse events and antibiotic resistance in Helicobacter pylori treatment. Scand J Gastroenterol Suppl. 1993;196:34–37. doi: 10.3109/00365529309098341. [DOI] [PubMed] [Google Scholar]
  16. Rauws E. A., Tytgat G. N. Cure of duodenal ulcer associated with eradication of Helicobacter pylori. Lancet. 1990 May 26;335(8700):1233–1235. doi: 10.1016/0140-6736(90)91301-p. [DOI] [PubMed] [Google Scholar]
  17. Tytgat G. N. Review article: treatments that impact favourably upon the eradication of Helicobacter pylori and ulcer recurrence. Aliment Pharmacol Ther. 1994 Aug;8(4):359–368. doi: 10.1111/j.1365-2036.1994.tb00303.x. [DOI] [PubMed] [Google Scholar]
  18. Ursi J. P., Ursi D., Ieven M., Pattyn S. R. Utility of an internal control for the polymerase chain reaction. Application to detection of Mycoplasma pneumoniae in clinical specimens. APMIS. 1992 Jul;100(7):635–639. doi: 10.1111/j.1699-0463.1992.tb03978.x. [DOI] [PubMed] [Google Scholar]
  19. Valentine J. L., Arthur R. R., Mobley H. L., Dick J. D. Detection of Helicobacter pylori by using the polymerase chain reaction. J Clin Microbiol. 1991 Apr;29(4):689–695. doi: 10.1128/jcm.29.4.689-695.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wagner S., Gebel M., Manns M. Therapie der Helicobacter pylori-Infektion: Aktueller Stand. Z Gastroenterol. 1993 Jul-Aug;31(7-8):459–463. [PubMed] [Google Scholar]
  21. Weiss J., Mecca J., da Silva E., Gassner D. Comparison of PCR and other diagnostic techniques for detection of Helicobacter pylori infection in dyspeptic patients. J Clin Microbiol. 1994 Jul;32(7):1663–1668. doi: 10.1128/jcm.32.7.1663-1668.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Xia H. X., Keane C. T., O'Morain C. A. Pre-formed urease activity of Helicobacter pylori as determined by a viable cell count technique--clinical implications. J Med Microbiol. 1994 Jun;40(6):435–439. doi: 10.1099/00222615-40-6-435. [DOI] [PubMed] [Google Scholar]
  23. von Wulffen H., Heesemann J., Bützow G. H., Löning T., Laufs R. Detection of Campylobacter pyloridis in patients with antrum gastritis and peptic ulcers by culture, complement fixation test, and immunoblot. J Clin Microbiol. 1986 Nov;24(5):716–720. doi: 10.1128/jcm.24.5.716-720.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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