Abstract
The detection of Helicobacter pylori in clinical and environmental samples by PCR sometimes requires removal of polymerase inhibitors. We have used a magnetic immunoseparation technique as pre-PCR treatment to facilitate direct detection of H. pylori in stool and water specimens. Rabbit hyperimmune antiserum was produced and magnetic beads were coated with purified immunoglobulin G, which reacted with and bound to both coccoid and rod-shaped forms of H. pylori. When PCR was applied for the detection of H. pylori from cultured samples, the number of organisms that was required for positive scores varied significantly. For a 3-day culture of H. pylori, samples containing 10(2) bacteria per ml are needed for a positive score; for a 6-day culture, samples containing 10(4) bacteria per ml are needed; and for a 10-day culture, samples containing 10(6) bacteria per ml are needed. These results indicate that the coccoid forms of H. pylori may have a different antigenicity and DNA content and are therefore more difficult to detect by immunomagnetic separation and PCR than the rod-shaped forms. Spiked samples with the addition of feces, spiked water samples, and a patient stool specimen were all scored positive with this technique.
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- Bickley J., Owen R. J., Fraser A. G., Pounder R. E. Evaluation of the polymerase chain reaction for detecting the urease C gene of Helicobacter pylori in gastric biopsy samples and dental plaque. J Med Microbiol. 1993 Nov;39(5):338–344. doi: 10.1099/00222615-39-5-338. [DOI] [PubMed] [Google Scholar]
- Biswas B., Vemulapalli R., Dutta S. K. Detection of Ehrlichia risticii from feces of infected horses by immunomagnetic separation and PCR. J Clin Microbiol. 1994 Sep;32(9):2147–2151. doi: 10.1128/jcm.32.9.2147-2151.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bode G., Mauch F., Malfertheiner P. The coccoid forms of Helicobacter pylori. Criteria for their viability. Epidemiol Infect. 1993 Dec;111(3):483–490. doi: 10.1017/s0950268800057216. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Catrenich C. E., Makin K. M. Characterization of the morphologic conversion of Helicobacter pylori from bacillary to coccoid forms. Scand J Gastroenterol Suppl. 1991;181:58–64. [PubMed] [Google Scholar]
- Danielsson D., Blomberg B., Järnerot G., Kosunen T. U. Heterogeneity of Campylobacter pylori as demonstrated by co-agglutination testing with rabbit antibodies. Scand J Gastroenterol Suppl. 1988;142:58–63. [PubMed] [Google Scholar]
- Engstrand L., Gustavsson S., Jörgensen A., Schwan A., Scheynius A. Inoculation of barrier-born pigs with Helicobacter pylori: a useful animal model for gastritis type B. Infect Immun. 1990 Jun;58(6):1763–1768. doi: 10.1128/iai.58.6.1763-1768.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Evans D. G., Karjalainen T. K., Evans D. J., Jr, Graham D. Y., Lee C. H. Cloning, nucleotide sequence, and expression of a gene encoding an adhesin subunit protein of Helicobacter pylori. J Bacteriol. 1993 Feb;175(3):674–683. doi: 10.1128/jb.175.3.674-683.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ferguson D. A., Jr, Li C., Patel N. R., Mayberry W. R., Chi D. S., Thomas E. Isolation of Helicobacter pylori from saliva. J Clin Microbiol. 1993 Oct;31(10):2802–2804. doi: 10.1128/jcm.31.10.2802-2804.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Handt L. K., Fox J. G., Dewhirst F. E., Fraser G. J., Paster B. J., Yan L. L., Rozmiarek H., Rufo R., Stalis I. H. Helicobacter pylori isolated from the domestic cat: public health implications. Infect Immun. 1994 Jun;62(6):2367–2374. doi: 10.1128/iai.62.6.2367-2374.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hazell S. L., Mitchell H. M., Hedges M., Shi X., Hu P. J., Li Y. Y., Lee A., Reiss-Levy E. Hepatitis A and evidence against the community dissemination of Helicobacter pylori via feces. J Infect Dis. 1994 Sep;170(3):686–689. doi: 10.1093/infdis/170.3.686. [DOI] [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]
- 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]
- Moshkowitz M., Gorea A., Arber N., Konikoff F., Berger S., Gilat T. Morphological transformation of Helicobacter pylori during prolonged incubation: association with decreased acid resistance. J Clin Pathol. 1994 Feb;47(2):172–174. doi: 10.1136/jcp.47.2.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Muir P., Nicholson F., Jhetam M., Neogi S., Banatvala J. E. Rapid diagnosis of enterovirus infection by magnetic bead extraction and polymerase chain reaction detection of enterovirus RNA in clinical specimens. J Clin Microbiol. 1993 Jan;31(1):31–38. doi: 10.1128/jcm.31.1.31-38.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Nilius M., Ströhle A., Bode G., Malfertheiner P. Coccoid like forms (CLF) of Helicobacter pylori. Enzyme activity and antigenicity. Zentralbl Bakteriol. 1993 Sep;280(1-2):259–272. doi: 10.1016/s0934-8840(11)80964-3. [DOI] [PubMed] [Google Scholar]
- Olsvik O., Popovic T., Skjerve E., Cudjoe K. S., Hornes E., Ugelstad J., Uhlén M. Magnetic separation techniques in diagnostic microbiology. Clin Microbiol Rev. 1994 Jan;7(1):43–54. doi: 10.1128/cmr.7.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seesod N., Lunderberg J., Hedrum A., Aslund L., Holder A., Thaithong S., Uhlén M. Immunomagnetic purification to facilitate DNA diagnosis of Plasmodium falciparum. J Clin Microbiol. 1993 Oct;31(10):2715–2719. doi: 10.1128/jcm.31.10.2715-2719.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas J. E., Gibson G. R., Darboe M. K., Dale A., Weaver L. T. Isolation of Helicobacter pylori from human faeces. Lancet. 1992 Nov 14;340(8829):1194–1195. doi: 10.1016/0140-6736(92)92894-l. [DOI] [PubMed] [Google Scholar]
- West A. P., Millar M. R., Tompkins D. S. Survival of Helicobacter pylori in water and saline. J Clin Pathol. 1990 Jul;43(7):609–609. doi: 10.1136/jcp.43.7.609-b. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wright D. J., Chapman P. A., Siddons C. A. Immunomagnetic separation as a sensitive method for isolating Escherichia coli O157 from food samples. Epidemiol Infect. 1994 Aug;113(1):31–39. doi: 10.1017/s0950268800051438. [DOI] [PMC free article] [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]