Nucleic Acids Research, 1992, Vol. 20, No. 19 5137-5142
© 1992
MOLECULAR BIOLOGY |
DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting
Department of Molecular Microbiology, Campus Box 8230, Washington University Medical School St Louis, MO 63110, USA 1Division of Infectious Diseases and Immunology, St Louis University School of Medicine St Louis, MO 63104, USA 2Department of Biology, Washington University St Louis, MO 63130, USA
* To whom correspondence should be addressed
Received June 16, 1992. Revised September 1, 1992. Accepted September 1, 1992.
The RAPD (or AP-PCR) DNA fingerprinting method was used to distinguish among clinical isolates of Helicobacter pylori, a bacterium whose long term carriage is associated with gastritis, peptic ulcers and gastric carcinomas. This method uses arbitrarily chosen oligonucleotides to prime DNA synthesis from genomlc sites to which they are fortuitously matched, or almost matched. Most 10-nt primers with ss 60% G + C yielded strain-specific arrays of up to 15 prominent fragments, as did most longer (
17-nt) primers, whereas most 10-nt primers with 50% G + C did not. Each of 64 Independent H.pylori isolates, 60 of which were from patients in the same hospital, was distinguishable with a single RAPD primer, which suggests a high level of DNA sequence diversity within this species. In contrast, isolates from initial and followup biopsies were indistinguishable in each of three cases tested.
+ Permanent address: USDA-ARS Plant Genetic Resources Unit, Cornell University, Geneva, NY 14456-0462, USA
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