Genomic DNA fingerprinting of clinical isolates of Helicobacter pylori using short oligonucleotide probes containing repetitive sequences.
The ability of oligonucleotide probes containing short repetitive sequence motifs to differentiate between isolates of Helicobacter pylori was investigated. Genomic DNA preparations from H. pylori were digested with the restriction enzyme HindIII, electrophoresed in agarose gels and transferred to nylon filters. Five separate oligonucleotide probes were tested for hybridization sequentially to fingerprint the digested DNA from a panel of 29 clinical isolates and one type strain of H. pylori, and their relative discriminatory abilities were assessed. Four probes, (GACA)4, (GT)8, (GTG)5 and (GGAT)4, were each shown to yield highly informative hybridization band profiles allowing differentiation of H. pylori isolates. The DNA fingerprints of individual isolates obtained with each probe were distinct and reproducible. Direct comparison with ribotyping revealed that oligonucleotide fingerprinting had far superior discriminatory power. Computer-assisted similarity analysis of (GGAT)4-generated hybridization profiles of pairwise combinations of H. pylori isolates revealed that there was no correlation between ribotype and oligonucleotide fingerprint patterns. The results of this study demonstrate that oligonucleotide probes containing microsatellite sequences provide a new and powerful tool for isolate discrimination of H. pylori.
Marshall
D G
DG
Moyne Institute of Preventive Medicine, Department of Microbiology, School of Dental Science, Trinity College, University of Dublin, Republic of Ireland. dmrshall@mail.tcd.i.e
Coleman
D C
DC
Sullivan
D J
DJ
Xia
H
H
O'Moráin
C A
CA
Smyth
C J
CJ
D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis*; D016680:Genome, Bacterial; D016480:Helicobacter pylori / Q000235:genetics* / Q000302:isolation & purification; D018895:Microsatellite Repeats; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D015203:Reproducibility of Results
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes