New Rapid Polymerase Chain Reaction-Immunochromatographic Assay for Porphyromonas gingivalis
- Autores: Kazuko Takada, Yoshiaki Sakaguchi, Chitoshi Oka, Masatomo Hirasawa
- Localización: Journal of periodontology, ISSN 0022-3492, Vol. 76, Nº. 4, 2005, págs. 508-512
- Idioma: inglés
- Enlaces
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Resumen
Background: A simple and rapid method for Porphyromonas gingivalis detection in clinical samples has been developed using polymerase chain reaction (PCR) and an immunochromatographic assay (ICA) with a lateral-flow device (strip) to detect species-specific 16S rRNA genes.
Methods: The PCR used a pair of primer sets labeled with fluorescein isothiocyanate (FITC) or biotin at each 5' terminus. The strip used a nitrocellulose membrane containing streptavidin conjugated to gold particles and anti-FITC line.
Results: PCR and ICA detected as few as 1 and 10 cells of P. gingivalis, respectively. ICA required 5 to 10 minutes more than the initial PCR. The amplifications were not observed in other oral black-pigmented bacteria at concentrations of 106 colony forming unit (CFU). The ICA strips showed bands at more than 104 CFU/ml equivalents in clinical samples from periodontitis.
Conclusions: A diagnostic assay based on PCR-ICA was developed for the detection of P. gingivalis, and results were obtained visually in 3 hours. PCR-ICA will be a valuable tool for the rapid detection of target bacteria by chair side. J Periodontol 2005;76:508-512.
