Abstract
The PCR primer set Hp1-Hp2, which amplifies a 109-bp fragment of the 16S rRNA gene of Helicobacter pylori, has been widely used for the detection of H. pylori in clinical specimens. We have examined 34 stool samples and 50 human tissue samples from H. pylori-infected and uninfected patients, five human leukocyte samples, and one human cell line by this PCR method. All of these specimens produced a 109-bp PCR product. When Escherichia coli DNA was used as the template, several nonspecific bands, but not the 109-bp band, were observed. No PCR products were generated when DNA samples from five different fungi were used as templates. These results indicate that this 109-bp PCR product was amplified from the human genome. The 109-bp PCR product generated from various clinical specimens also hybridized with the probe pHp, corresponding to a region internal to the PCR product of Hp1-Hp2. We conclude that the 16S rRNA gene PCR with the primer set Hp1-Hp2 is not specific and cannot be used to detect H. pylori in clinical specimens.
Full Text
The Full Text of this article is available as a PDF (372.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Chong S. K., Lou Q., Asnicar M. A., Zimmerman S. E., Croffie J. M., Lee C. H., Fitzgerald J. F. Helicobacter pylori infection in recurrent abdominal pain in childhood: comparison of diagnostic tests and therapy. Pediatrics. 1995 Aug;96(2 Pt 1):211–215. [PubMed] [Google Scholar]
- 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]
- Lee C. H., Hu S. T., Swiatek P. J., Moseley S. L., Allen S. D., So M. Isolation of a novel transposon which carries the Escherichia coli enterotoxin STII gene. J Bacteriol. 1985 May;162(2):615–620. doi: 10.1128/jb.162.2.615-620.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee C. H., Lu J. J., Bartlett M. S., Durkin M. M., Liu T. H., Wang J., Jiang B., Smith J. W. Nucleotide sequence variation in Pneumocystis carinii strains that infect humans. J Clin Microbiol. 1993 Mar;31(3):754–757. doi: 10.1128/jcm.31.3.754-757.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Makimura K., Murayama S. Y., Yamaguchi H. Detection of a wide range of medically important fungi by the polymerase chain reaction. J Med Microbiol. 1994 May;40(5):358–364. doi: 10.1099/00222615-40-5-358. [DOI] [PubMed] [Google Scholar]
- 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]
- Mapstone N. P., Lynch D. A., Lewis F. A., Axon A. T., Tompkins D. S., Dixon M. F., Quirke P. PCR identification of Helicobacter pylori in faeces from gastritis patients. Lancet. 1993 Feb 13;341(8842):447–447. doi: 10.1016/0140-6736(93)93053-4. [DOI] [PubMed] [Google Scholar]
- Marshall B. J., Warren J. R., Francis G. J., Langton S. R., Goodwin C. S., Blincow E. D. Rapid urease test in the management of Campylobacter pyloridis-associated gastritis. Am J Gastroenterol. 1987 Mar;82(3):200–210. [PubMed] [Google Scholar]
- Morera-Brenes B., Sierra R., Barrantes R., Jonasson J., Nord C. E. Helicobacter pylori in a Costa Rican dyspeptic patient population. Eur J Clin Microbiol Infect Dis. 1994 Mar;13(3):253–257. doi: 10.1007/BF01974546. [DOI] [PubMed] [Google Scholar]
- Roosendaal R., Kuipers E. J., van den Brule A. J., Peña A. S., Uyterlinde A. M., Walboomers J. M., Meuwissen S. G., de Graaff J. Importance of the fiberoptic endoscope cleaning procedure for detection of Helicobacter pylori in gastric biopsy specimens by PCR. J Clin Microbiol. 1994 Apr;32(4):1123–1126. doi: 10.1128/jcm.32.4.1123-1126.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]