Quantitation of Helicobacter pylori in dental plaque samples by competitive polymerase chain reaction
Open Access
- 1 March 2000
- journal article
- Published by BMJ in Journal of Clinical Pathology
- Vol. 53 (3), 218-222
- https://doi.org/10.1136/jcp.53.3.218
Abstract
Aim—To establish a competitive PCR (cPCR) assay for quantitation of H pylori organisms in dental plaque samples. Methods—The cPCR coamplified target H pylori DNA and a known amount of internal standard template in the same tube with the same primers directed to 0.86 kb DNA of H pylori. The internal standard was a synthesised DNA bearing the same primer recognition sites at two ends and a non-homologous core sequence as the target DNA fragment. Quantitation was based on determination of the relative, not absolute, amounts of the differently sized and [32P]-dCTP labelled products derived from H pylori DNA and the competitive internal standard after gel electrophoresis separation. Results—A significant correlation between known amounts of H pylori added to dental plaque samples and the results of the cPCR was found, and a standard line was developed which allowed quantitation of H pylori in the plaque samples. cPCR was performed on supragingival plaque samples from 10 adult patients with H pylori infection in the stomach, and from five adults and six children without H pylori infection in the stomach. The ranges of H pylori numbers were 1–213 (median 25), 6–76 (10), and 4–94 (14) cells/mg of dental plaque in the three groups, respectively. Conclusions—cPCR is useful for quantitation of H pylori in supragingival dental plaque samples; however, the number of the organisms in dental plaque samples seems very low.Keywords
This publication has 24 references indexed in Scilit:
- A newly developed PCR assay ofH. pylori in gastric biopsy, saliva, and fecesDigestive Diseases and Sciences, 1996
- Role of Dental Plaque in the Transmission of Helicobacter Pylori InfectionJournal of Clinical Gastroenterology, 1996
- Helicobacter pylori in the mouth - negative isolation from dental plaque and salivaEuropean Journal of Gastroenterology & Hepatology, 1996
- High prevalence of Helicobacter pylori in saliva demonstrated by a novel PCR assay.Journal of Clinical Pathology, 1995
- Development of a PCR-based technique for detection ofHelicobacter pyloriFEMS Immunology & Medical Microbiology, 1995
- Evaluation of the polymerase chain reaction for detecting the urease C gene of Helicobacter pylori in gastric biopsy samples and dental plaqueJournal of Medical Microbiology, 1993
- Identification of Helicobacter pylori DNA in the mouths and stomachs of patients with gastritis using PCR.Journal of Clinical Pathology, 1993
- H pylori in dental plaquesThe Lancet, 1993
- Absence ofHelicobacter pylori in dental plaques inHelicobacter pylori positive dyspeptic patientsEuropean Journal of Clinical Microbiology & Infectious Diseases, 1993
- Helicobacter pylori in dental plaqueThe Lancet, 1993