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. 1995 May;33(5):1344–1347. doi: 10.1128/jcm.33.5.1344-1347.1995

Diagnostic assay for Helicobacter hepaticus based on nucleotide sequence of its 16S rRNA gene.

J K Battles 1, J C Williamson 1, K M Pike 1, P L Gorelick 1, J M Ward 1, M A Gonda 1
PMCID: PMC228160  PMID: 7542270

Abstract

Conserved primers were used to PCR amplify 95% of the Helicobacter hepaticus 16S rRNA gene. Its sequence was determined and aligned to those of related bacteria, enabling the selection of primers to highly diverged regions of the 16S rRNA gene and an oligonucleotide probe for the development of a PCR-liquid hybridization assay. This assay was shown to be both sensitive and specific for H. hepaticus 16S rRNA gene sequences.

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Selected References

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  1. Blaser M. J. Gastric Campylobacter-like organisms, gastritis, and peptic ulcer disease. Gastroenterology. 1987 Aug;93(2):371–383. doi: 10.1016/0016-5085(87)91028-6. [DOI] [PubMed] [Google Scholar]
  2. Bronsdon M. A., Goodwin C. S., Sly L. I., Chilvers T., Schoenknecht F. D. Helicobacter nemestrinae sp. nov., a spiral bacterium found in the stomach of a pigtailed macaque (Macaca nemestrina) Int J Syst Bacteriol. 1991 Jan;41(1):148–153. doi: 10.1099/00207713-41-1-148. [DOI] [PubMed] [Google Scholar]
  3. Desai M., Linton D., Owen R. J., Cameron H., Stanley J. Genetic diversity of Helicobacter pylori indexed with respect to clinical symptomatology, using a 16S rRNA and a species-specific DNA probe. J Appl Bacteriol. 1993 Dec;75(6):574–582. doi: 10.1111/j.1365-2672.1993.tb01597.x. [DOI] [PubMed] [Google Scholar]
  4. Edwards U., Rogall T., Blöcker H., Emde M., Böttger E. C. Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res. 1989 Oct 11;17(19):7843–7853. doi: 10.1093/nar/17.19.7843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Engstrand L., Nguyen A. M., Graham D. Y., el-Zaatari F. A. Reverse transcription and polymerase chain reaction amplification of rRNA for detection of Helicobacter species. J Clin Microbiol. 1992 Sep;30(9):2295–2301. doi: 10.1128/jcm.30.9.2295-2301.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fennell C. L., Totten P. A., Quinn T. C., Patton D. L., Holmes K. K., Stamm W. E. Characterization of Campylobacter-like organisms isolated from homosexual men. J Infect Dis. 1984 Jan;149(1):58–66. doi: 10.1093/infdis/149.1.58. [DOI] [PubMed] [Google Scholar]
  7. Fox G. E., Stackebrandt E., Hespell R. B., Gibson J., Maniloff J., Dyer T. A., Wolfe R. S., Balch W. E., Tanner R. S., Magrum L. J. The phylogeny of prokaryotes. Science. 1980 Jul 25;209(4455):457–463. doi: 10.1126/science.6771870. [DOI] [PubMed] [Google Scholar]
  8. Fox J. G., Dewhirst F. E., Tully J. G., Paster B. J., Yan L., Taylor N. S., Collins M. J., Jr, Gorelick P. L., Ward J. M. Helicobacter hepaticus sp. nov., a microaerophilic bacterium isolated from livers and intestinal mucosal scrapings from mice. J Clin Microbiol. 1994 May;32(5):1238–1245. doi: 10.1128/jcm.32.5.1238-1245.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fox J. G., Edrise B. M., Cabot E. B., Beaucage C., Murphy J. C., Prostak K. S. Campylobacter-like organisms isolated from gastric mucosa of ferrets. Am J Vet Res. 1986 Feb;47(2):236–239. [PubMed] [Google Scholar]
  10. Fox J. G., Paster B. J., Dewhirst F. E., Taylor N. S., Yan L. L., Macuch P. J., Chmura L. M. Helicobacter mustelae isolation from feces of ferrets: evidence to support fecal-oral transmission of a gastric Helicobacter. Infect Immun. 1992 Feb;60(2):606–611. doi: 10.1128/iai.60.2.606-611.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gray M. W., Sankoff D., Cedergren R. J. On the evolutionary descent of organisms and organelles: a global phylogeny based on a highly conserved structural core in small subunit ribosomal RNA. Nucleic Acids Res. 1984 Jul 25;12(14):5837–5852. doi: 10.1093/nar/12.14.5837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greisen K., Loeffelholz M., Purohit A., Leong D. PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid. J Clin Microbiol. 1994 Feb;32(2):335–351. doi: 10.1128/jcm.32.2.335-351.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Hoshina S., Kahn S. M., Jiang W., Green P. H., Neu H. C., Chin N., Morotomi M., LoGerfo P., Weinstein I. B. Direct detection and amplification of Helicobacter pylori ribosomal 16S gene segments from gastric endoscopic biopsies. Diagn Microbiol Infect Dis. 1990 Nov-Dec;13(6):473–479. doi: 10.1016/0732-8893(90)90079-b. [DOI] [PubMed] [Google Scholar]
  15. Lane D. J., Pace B., Olsen G. J., Stahl D. A., Sogin M. L., Pace N. R. Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6955–6959. doi: 10.1073/pnas.82.20.6955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lee A., Hazell S. L., O'Rourke J., Kouprach S. Isolation of a spiral-shaped bacterium from the cat stomach. Infect Immun. 1988 Nov;56(11):2843–2850. doi: 10.1128/iai.56.11.2843-2850.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Mapstone N. P., Lynch D. A., Lewis F. A., Axon A. T., Tompkins D. S., Dixon M. F., Quirke P. Identification of Helicobacter pylori DNA in the mouths and stomachs of patients with gastritis using PCR. J Clin Pathol. 1993 Jun;46(6):540–543. doi: 10.1136/jcp.46.6.540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marshall B. J., Warren J. R. Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet. 1984 Jun 16;1(8390):1311–1315. doi: 10.1016/s0140-6736(84)91816-6. [DOI] [PubMed] [Google Scholar]
  20. Morotomi M., Hoshina S., Green P., Neu H. C., LoGerfo P., Watanabe I., Mutai M., Weinstein I. B. Oligonucleotide probe for detection and identification of Campylobacter pylori. J Clin Microbiol. 1989 Dec;27(12):2652–2655. doi: 10.1128/jcm.27.12.2652-2655.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nguyen A. M., Engstrand L., Genta R. M., Graham D. Y., el-Zaatari F. A. Detection of Helicobacter pylori in dental plaque by reverse transcription-polymerase chain reaction. J Clin Microbiol. 1993 Apr;31(4):783–787. doi: 10.1128/jcm.31.4.783-787.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Olsen G. J., Lane D. J., Giovannoni S. J., Pace N. R., Stahl D. A. Microbial ecology and evolution: a ribosomal RNA approach. Annu Rev Microbiol. 1986;40:337–365. doi: 10.1146/annurev.mi.40.100186.002005. [DOI] [PubMed] [Google Scholar]
  23. Pace N. R., Olsen G. J., Woese C. R. Ribosomal RNA phylogeny and the primary lines of evolutionary descent. Cell. 1986 May 9;45(3):325–326. doi: 10.1016/0092-8674(86)90315-6. [DOI] [PubMed] [Google Scholar]
  24. Paster B. J., Lee A., Fox J. G., Dewhirst F. E., Tordoff L. A., Fraser G. J., O'Rourke J. L., Taylor N. S., Ferrero R. Phylogeny of Helicobacter felis sp. nov., Helicobacter mustelae, and related bacteria. Int J Syst Bacteriol. 1991 Jan;41(1):31–38. doi: 10.1099/00207713-41-1-31. [DOI] [PubMed] [Google Scholar]
  25. Romaniuk P. J., Trust T. J. Rapid identification of Campylobacter species using oligonucleotide probes to 16S ribosomal RNA. Mol Cell Probes. 1989 Jun;3(2):133–142. doi: 10.1016/0890-8508(89)90024-8. [DOI] [PubMed] [Google Scholar]
  26. Romaniuk P. J., Zoltowska B., Trust T. J., Lane D. J., Olsen G. J., Pace N. R., Stahl D. A. Campylobacter pylori, the spiral bacterium associated with human gastritis, is not a true Campylobacter sp. J Bacteriol. 1987 May;169(5):2137–2141. doi: 10.1128/jb.169.5.2137-2141.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schauer D. B., Ghori N., Falkow S. Isolation and characterization of "Flexispira rappini" from laboratory mice. J Clin Microbiol. 1993 Oct;31(10):2709–2714. doi: 10.1128/jcm.31.10.2709-2714.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Solnick J. V., O'Rourke J., Lee A., Paster B. J., Dewhirst F. E., Tompkins L. S. An uncultured gastric spiral organism is a newly identified Helicobacter in humans. J Infect Dis. 1993 Aug;168(2):379–385. doi: 10.1093/infdis/168.2.379. [DOI] [PubMed] [Google Scholar]
  29. Stanley J., Burnens A. P., Linton D., On S. L., Costas M., Owen R. J. Campylobacter helveticus sp. nov., a new thermophilic species from domestic animals: characterization, and cloning of a species-specific DNA probe. J Gen Microbiol. 1992 Nov;138(11):2293–2303. doi: 10.1099/00221287-138-11-2293. [DOI] [PubMed] [Google Scholar]
  30. Stanley J., Linton D., Burnens A. P., Dewhirst F. E., Owen R. J., Porter A., On S. L., Costas M. Helicobacter canis sp. nov., a new species from dogs: an integrated study of phenotype and genotype. J Gen Microbiol. 1993 Oct;139(10):2495–2504. doi: 10.1099/00221287-139-10-2495. [DOI] [PubMed] [Google Scholar]
  31. Taylor D. E., Eaton M., Chang N., Salama S. M. Construction of a Helicobacter pylori genome map and demonstration of diversity at the genome level. J Bacteriol. 1992 Nov;174(21):6800–6806. doi: 10.1128/jb.174.21.6800-6806.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Totten P. A., Fennell C. L., Tenover F. C., Wezenberg J. M., Perine P. L., Stamm W. E., Holmes K. K. Campylobacter cinaedi (sp. nov.) and Campylobacter fennelliae (sp. nov.): two new Campylobacter species associated with enteric disease in homosexual men. J Infect Dis. 1985 Jan;151(1):131–139. doi: 10.1093/infdis/151.1.131. [DOI] [PubMed] [Google Scholar]
  33. Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet. 1983 Jun 4;1(8336):1273–1275. [PubMed] [Google Scholar]
  34. Vandamme P., Falsen E., Rossau R., Hoste B., Segers P., Tytgat R., De Ley J. Revision of Campylobacter, Helicobacter, and Wolinella taxonomy: emendation of generic descriptions and proposal of Arcobacter gen. nov. Int J Syst Bacteriol. 1991 Jan;41(1):88–103. doi: 10.1099/00207713-41-1-88. [DOI] [PubMed] [Google Scholar]
  35. Ward J. M., Anver M. R., Haines D. C., Benveniste R. E. Chronic active hepatitis in mice caused by Helicobacter hepaticus. Am J Pathol. 1994 Oct;145(4):959–968. [PMC free article] [PubMed] [Google Scholar]
  36. Ward J. M., Fox J. G., Anver M. R., Haines D. C., George C. V., Collins M. J., Jr, Gorelick P. L., Nagashima K., Gonda M. A., Gilden R. V. 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 Aug 17;86(16):1222–1227. doi: 10.1093/jnci/86.16.1222. [DOI] [PubMed] [Google Scholar]
  37. Weisburg W. G., Barns S. M., Pelletier D. A., Lane D. J. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991 Jan;173(2):697–703. doi: 10.1128/jb.173.2.697-703.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. van Zwet A. A., Thijs J. C., Kooistra-Smid A. M., Schirm J., Snijder J. A. Use of PCR with feces for detection of Helicobacter pylori infections in patients. J Clin Microbiol. 1994 May;32(5):1346–1348. doi: 10.1128/jcm.32.5.1346-1348.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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