PMID: 8602346Dec 1, 1995Paper

Colorimetric microtiter plate based assay for detection and quantification of amplified Actinobacillus actinomycetemcomitans DNA

Oral Microbiology and Immunology
O FujiseK Maeda

Abstract

We developed a colorimetric microtiter plate-based assay for the detection and quantification of polymerase chain reaction-amplified DNA fragment specific for Actinobacillus actinomycetemcomitans. We amplified the 396-bp leukotoxin-specific DNA fragment by using two oligonucleotide primers, one carrying a biotin group at the 5' end and another one with a digoxigenin at the 5' end. Following amplification, the biotinylated polymerase chain reaction products were applied to a microtiter well precoated with avidin. The colorimetric detection and quantification were achieved by an enzyme-linked immunosorbent assay using alkaline phosphatase-conjugated anti-digoxigenin antibody. The detection limit of the colorimetric assay was found to be as little as 500 fg of purified A. actinomycetemcomitans DNA and as few as 50 A. actinomycetemcomitans. Therefore, this colorimetric assay was able to estimate the amount of A. actinomycetemcomitans in subgingival plaque samples. We concluded that the colorimetric assay of the PCR product is a very useful method not only to detect the presence of A. actinomycetemcomitans but also to quantify the amount of A. actinomycetemcomitans in large numbers of subgingival plaque samples.

References

Apr 1, 1992·Journal of Periodontology·S S Socransky, A D Haffajee
Jan 1, 1985·Journal of Clinical Periodontology·J J Zambon
Jun 1, 1994·Oral Microbiology and Immunology·A BodinkaH Karch
May 1, 1994·Journal of Periodontology·L WolffD Aeppli
May 1, 1994·Journal of Clinical Microbiology·E J LeysP A Fuerst
Mar 1, 1993·Journal of Clinical Microbiology·G M van der VlietP R Klatser
Jun 1, 1994·Periodontology 2000·A D Haffajee, S S Socransky
May 1, 1994·Journal of Periodontology·Larry WolffDorothee Aeppli

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Citations

Feb 9, 2008·Critical Reviews in Microbiology·Simonetta D'ErcoleRaffaele Piccolomini
Nov 25, 2004·Journal of Clinical Periodontology·Juan Manuel MorilloAugusto Silva

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