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New Rapid Polymerase Chain Reaction‐Immunochromatographic Assay for Porphyromonas gingivalis
Author(s) -
Takada Kazuko,
Sakaguchi Yoshiaki,
Oka Chitoshi,
Hirasawa Masatomo
Publication year - 2005
Publication title -
journal of periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.036
H-Index - 156
eISSN - 1943-3670
pISSN - 0022-3492
DOI - 10.1902/jop.2005.76.4.508
Subject(s) - porphyromonas gingivalis , polymerase chain reaction , microbiology and biotechnology , nitrocellulose , primer (cosmetics) , 16s ribosomal rna , fluorescein isothiocyanate , bacteria , chemistry , real time polymerase chain reaction , streptavidin , chromatography , biology , biotin , fluorescence , gene , membrane , biochemistry , genetics , physics , organic chemistry , quantum mechanics
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 10 6 colony forming unit (CFU). The ICA strips showed bands at more than 10 4 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 .