Title
Comparison of viable plate count, turbidity measurement and real-time PCR for quantification of Porphyromonas gingivalis Comparison of viable plate count, turbidity measurement and real-time PCR for quantification of Porphyromonas gingivalis
Author
Faculty/Department
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences. Pharmacy
Publication type
article
Publication
Oxford ,
Subject
Biology
Human medicine
Engineering sciences. Technology
Source (journal)
Letters in applied microbiology. - Oxford
Volume/pages
60(2015) :1 , p. 79-84
ISSN
0266-8254
ISI
000346590200012
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
The viable plate count (VPC) is considered as the reference method for bacterial enumeration in periodontal microbiology but shows some important limitations for anaerobic bacteria. As anaerobes such as Porphyromonas gingivalis are difficult to culture, VPC becomes time-consuming and less sensitive. Hence, efficient normalization of experimental data to bacterial cell count requires alternative rapid and reliable quantification methods. This study compared the performance of VPC with that of turbidity measurement and real-time PCR (qPCR) in an experimental context using highly concentrated bacterial suspensions. Our TaqMan-based qPCR assay for P. gingivalis 16S rRNA proved to be sensitive and specific. Turbidity measurements offer a fast method to assess P. gingivalis growth, but suffer from high variability and a limited dynamic range. VPC was very time-consuming and less repeatable than qPCR. Our study concludes that qPCR provides the most rapid and precise approach for P. gingivalis quantification. Although our data were gathered in a specific research context, we believe that our conclusions on the inferior performance of VPC and turbidity measurements in comparison to qPCR can be extended to other research and clinical settings and even to other difficult-to-culture micro-organisms.
E-info
https://repository.uantwerpen.be/docman/iruaauth/0945e0/97602831aec.pdf
Full text (open access)
https://repository.uantwerpen.be/docman/irua/3534ab/9094.pdf
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