Abstract
Quantitative PCR (qPCR) has recently been used to quantify microorganisms in complex communities, including dental plaque biofilms. However, there is variability in the qPCR protocols being used. This study was designed to evaluate the validity of two of these variables with the intent of developing a more standardized qPCR protocol. The two variables evaluated were (1) the use of DNA content versus actual cell counts to estimate bacterial numbers in mixed plaque samples and (2) the effectiveness of three different universal primers versus species specific primers in amplifying specific target pathogens in these samples. Results lead to the development of a standardized protocol that was shown to be highly reproducible as demonstrated by low coefficients of variation. The results also confirmed that this standardized qPCR protocol can be used as a sensitive method for quantifying specific bacterial species in human plaque samples.
Document Type
Article
Publication Date
2008
Digital Object Identifier (DOI)
http://dx.doi.org/10.2174/1874210600802010049
Repository Citation
Kirakodu, Sreenatha S.; Govindaswami, M.; Novak, Michael John; Ebersole, Jeffrey L.; and Novak, Karen F., "Optimizing qPCR for the Quantification of Periodontal Pathogens in a Complex Plaque Biofilm" (2008). Center for Oral Health Research Faculty Publications. 2.
https://uknowledge.uky.edu/cohr_facpub/2
Included in
Genetics Commons, Immunology of Infectious Disease Commons, Molecular Genetics Commons, Oral Biology and Oral Pathology Commons
Notes/Citation Information
Published in The Open Dentistry Journal, v. 2, p. 49-55.
© Kirakodu et al. ; Licensee Bentham Open .
This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/ by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.