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Titolo:
A sensitive method for detecting Porphyromonas gingivalis by polymerase chain reaction and its possible clinical application
Autore:
Nozaki, T; Kusumoto, Y; Kitamura, M; Hirano, H; Kohyama, A; Hayakawa, M; Takiguchi, H; Abiko, Y; Murakami, S; Okada, H;
Indirizzi:
Osaka Univ, Dept Periodontol, Div Oral Biol & Dis Control, Grad Sch dent, Suita, Osaka 5650871, Japan Osaka Univ Suita Osaka Japan 5650871 ch dent, Suita, Osaka 5650871, Japan Nihon Univ, Sch Dent, Dept Biochem, Matsudo, Chiba 271, Japan Nihon Univ Matsudo Chiba Japan 271 ept Biochem, Matsudo, Chiba 271, Japan
Titolo Testata:
JOURNAL OF PERIODONTOLOGY
fascicolo: 9, volume: 72, anno: 2001,
pagine: 1228 - 1235
SICI:
0022-3492(200109)72:9<1228:ASMFDP>2.0.ZU;2-Y
Fonte:
ISI
Lingua:
ENG
Soggetto:
ORAL PLAQUE SAMPLES; ACTINOBACILLUS-ACTINOMYCETEMCOMITANS; DNA PROBES; OLIGODEOXYNUCLEOTIDE PROBES; PERIODONTAL ATTACHMENT; BACTEROIDES-GINGIVALIS; RISK INDICATORS; BACTERIA; IDENTIFICATION; DIAGNOSIS;
Keywords:
immunofluorescence assay, indirect; periodontitis/diagnosis; periodontal pockets/microbiology; periodontitis/microbiology; polymerase chain reaction; Porphyromonas gingivalis; bacterial outer membrane proteins;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
30
Recensione:
Indirizzi per estratti:
Indirizzo: Nozaki, T Osaka Univ, Dept Periodontol, Div Oral Biol & Dis Control, Grad Sch dent, 1-8 Yamadaoka, Suita, Osaka 5650871, Japan Osaka Univ 1-8 Yamadaoka Suita Osaka Japan 5650871 650871, Japan
Citazione:
T. Nozaki et al., "A sensitive method for detecting Porphyromonas gingivalis by polymerase chain reaction and its possible clinical application", J PERIODONT, 72(9), 2001, pp. 1228-1235

Abstract

Background: It is useful for the clinical diagnosis of periodontitis to monitor the colonization of periodontopathic bacteria in periodontal pockets. In this study, we attempted to establish and possibly identify the clinical application of a sensitive method to detect Porphyromonas gingivalis (Pg.), one of the putative periodontopathic bacteria related to chronic periodontitis. Methods: Genomic DNA extracted from cultured Pg. 381 and clinically isolated subgingival plaque samples were used as a template of polymerase chain reaction (PCR). We designed primers to amplify the genomic DNA coding 40 kDaouter membrane protein (OMP), one of the unique proteins to all strains ofPg. The efficiency and specificity of amplification were evaluated by agarose gel electrophoresis and subsequent Southern hybridization with a digoxygenin-labeled oligonucleotide probe. Results: Fewer than 100 Pg. bacterial cells in the specimen were reproducibly detected by PCR-hybridization assay. This PCR-hybridization assay was at least 100 times more sensitive than the conventional indirect immunofluorescence assay (IIF). Furthermore, the imaging analysis showed that there isa linear correlation between the strength of the signal and the cell number of Pg. from which the template DNA was extracted semiquantitatively. It is noteworthy that the PCR assay could also be applied to detect Pg. from clinical plaque samples and that it was approximately 100 times more sensitive than a conventional IIF assay. Conclusion: The PCR assay established in this study can be a powerful toolto detect Pg. in periodontal pockets and monitor the colonization and/or recolonization of P.g. at the very early phase.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/09/20 alle ore 13:52:14