Identification of H. pylori in saliva by a nested PCR assay derived from a newly cloned DNA probe.
A novel probe was developed from genomic DNA of Helicobacter pylori ATCC type strain 43629. It hybridized with all 73 H. pylori clinical isolates tested but not with any of 183 non-H. pylori DNAs in dot blot hybridization. Typing tests revealed 41 different HaeIII-digestion patterns from 57 H. pylori strains tested. Based on the sequence of the probe, a nested PCR was developed that detected as little as 2 fg of H. pylori DNA or approximately equivalent to one cell. No PCR products were amplified from any of 21 non-H. pylori strains tested. Using this nested PCR, H. pylori DNA was detected in 33 of 45 (73%) saliva samples collected from patients with gastric H. pylori infection. These data suggest that the probe is useful for typing H. pylori and that the nested PCR is a valuable tool for detecting H. pylori DNA in saliva.
Jiang
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C
Department of Internal Medicine, James H. Quillen College of Medicine, East Tennessee State University, and Veterans Affairs Medical Center, Johnson City 37684, USA.
Li
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C
Ha
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T
Ferguson
D A
DA
Chi
D S
DS
Laffan
J J
JJ
Thomas
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E
D016172:DNA Fingerprinting; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis*; D005821:Genetic Techniques; D016680:Genome, Bacterial; D016480:Helicobacter pylori / Q000145:classification / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012463:Saliva / Q000382:microbiology*; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004269:DNA, Bacterial