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. 1997 Apr;35(4):995–998. doi: 10.1128/jcm.35.4.995-998.1997

Complex polysaccharides as PCR inhibitors in feces: Helicobacter pylori model.

L Monteiro 1, D Bonnemaison 1, A Vekris 1, K G Petry 1, J Bonnet 1, R Vidal 1, J Cabrita 1, F Mégraud 1
PMCID: PMC229720  PMID: 9157172

Abstract

A model was developed to study inhibitors present in feces which prevent the use of PCR for the detection of Helicobacter pylori. A DNA fragment amplified with the same primers as H. pylori was used to spike samples before extraction by a modified QIAamp tissue method. Inhibitors, separated on an Ultrogel AcA44 column, were characterized. Inhibitors in feces are complex polysaccharides possibly originating from vegetable material in the diet.

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Selected References

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  1. Brian M. J., Frosolono M., Murray B. E., Miranda A., Lopez E. L., Gomez H. F., Cleary T. G. Polymerase chain reaction for diagnosis of enterohemorrhagic Escherichia coli infection and hemolytic-uremic syndrome. J Clin Microbiol. 1992 Jul;30(7):1801–1806. doi: 10.1128/jcm.30.7.1801-1806.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Demeke T., Adams R. P. The effects of plant polysaccharides and buffer additives on PCR. Biotechniques. 1992 Mar;12(3):332–334. [PubMed] [Google Scholar]
  3. Do N., Adams R. P. A simple technique for removing plant polysaccharide contaminants from DNA. Biotechniques. 1991 Feb;10(2):162–166. [PubMed] [Google Scholar]
  4. Fang G., Hammar S., Grumet R. A quick and inexpensive method for removing polysaccharides from plant genomic DNA. Biotechniques. 1992 Jul;13(1):52-4, 56. [PubMed] [Google Scholar]
  5. Frankel G., Riley L., Giron J. A., Valmassoi J., Friedmann A., Strockbine N., Falkow S., Schoolnik G. K. Detection of Shigella in feces using DNA amplification. J Infect Dis. 1990 Jun;161(6):1252–1256. doi: 10.1093/infdis/161.6.1252. [DOI] [PubMed] [Google Scholar]
  6. Furukawa K., Bhavanandan V. P. Influences of anionic polysaccharides on DNA synthesis in isolated nuclei and by DNA polymerase alpha: correlation of observed effects with properties of the polysaccharides. Biochim Biophys Acta. 1983 Sep 9;740(4):466–475. doi: 10.1016/0167-4781(83)90096-9. [DOI] [PubMed] [Google Scholar]
  7. Hammar M., Tyszkiewicz T., Wadström T., O'Toole P. W. Rapid detection of Helicobacter pylori in gastric biopsy material by polymerase chain reaction. J Clin Microbiol. 1992 Jan;30(1):54–58. doi: 10.1128/jcm.30.1.54-58.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Islam D., Lindberg A. A. Detection of Shigella dysenteriae type 1 and Shigella flexneri in feces by immunomagnetic isolation and polymerase chain reaction. J Clin Microbiol. 1992 Nov;30(11):2801–2806. doi: 10.1128/jcm.30.11.2801-2806.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jiang X., Wang J., Graham D. Y., Estes M. K. Detection of Norwalk virus in stool by polymerase chain reaction. J Clin Microbiol. 1992 Oct;30(10):2529–2534. doi: 10.1128/jcm.30.10.2529-2534.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Liu C., Smith B. M., Ajito K., Komatsu H., Gomez-Paloma L., Li T., Theodorakis E. A., Nicolaou K. C., Vogt P. K. Sequence-selective carbohydrate-DNA interaction: dimeric and monomeric forms of the calicheamicin oligosaccharide interfere with transcription factor function. Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):940–944. doi: 10.1073/pnas.93.2.940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lund A., Wasteson Y., Olsvik O. Immunomagnetic separation and DNA hybridization for detection of enterotoxigenic Escherichia coli in a piglet model. J Clin Microbiol. 1991 Oct;29(10):2259–2262. doi: 10.1128/jcm.29.10.2259-2262.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Shirai H., Nishibuchi M., Ramamurthy T., Bhattacharya S. K., Pal S. C., Takeda Y. Polymerase chain reaction for detection of the cholera enterotoxin operon of Vibrio cholerae. J Clin Microbiol. 1991 Nov;29(11):2517–2521. doi: 10.1128/jcm.29.11.2517-2521.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Siebert P. D., Larrick J. W. PCR MIMICS: competitive DNA fragments for use as internal standards in quantitative PCR. Biotechniques. 1993 Feb;14(2):244–249. [PubMed] [Google Scholar]
  16. Walsh P. S., Metzger D. A., Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques. 1991 Apr;10(4):506–513. [PubMed] [Google Scholar]
  17. Wilde J., Eiden J., Yolken R. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol. 1990 Jun;28(6):1300–1307. doi: 10.1128/jcm.28.6.1300-1307.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. de Lamballerie X., Zandotti C., Vignoli C., Bollet C., de Micco P. A one-step microbial DNA extraction method using "Chelex 100" suitable for gene amplification. Res Microbiol. 1992 Oct;143(8):785–790. doi: 10.1016/0923-2508(92)90107-y. [DOI] [PubMed] [Google Scholar]
  19. 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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