Porphyromonas gingivalis: A Methodology for Culturing, Extracting, and Identifying Fimbriated Strains
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Authors
Jeppesen, Addison B.
Issue Date
2018-04-17
Volume
Issue
Type
Thesis
Language
en_US
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Abstract
Background/purpose – Fimbriated Porphyromonas gingivalis has been shown to contribute to periodontitis, atherosclerosis, and arthritis. While many studies have been done on the bacterium itself, there is little information on selectively culturing and identifying P. gingivalis from a sulcal swab. To further the work of establishing a link between the bacterium and these health conditions, a selective and repeatable method had to be established to detect the presence of fimbriated strains of P. gingivalis. The present study aimed to establish working protocols for identification of P. gingivalis variants in sulcal swabs and serve as the launching point for future studies looking into early diagnosis and prevention of periodontal disease.|Methods – Although several different methods were in the course of this study, only the most effective, selective, and repeatable processes were reported on herewith. Incubation of the paper points in a blood agar media was found to produce detectable levels of P. gingivalis only when incubated for at least 72 hours. Extraction was performed using PureLink® Genomic DNA Kits that used filters as opposed to magnetic beads, and the manufacturers protocols were followed for gram-negative bacteria. The extracted DNA was then equalized and prepared for PCR assay using the eight primers detailed in this study. Positive bands were recorded following gel electrophoresis and unanticipated results were verified through DNA sequencing.|Results – Through experimentation we were able to establish a successful means of detecting the presence of fimbriated P. gingivalis from sulcal swabs, and establish which primers are effective in binding to the desired genomes. Despite the efficacy of these protocols, alterations may be required based on the lab personnel’s needs and desired outcomes. |Conclusion – The present study establishes an effective method of culturing, extracting and identifying fimbriated strains of P. gingivalis from sulcal swabs and frozen samples. Future work in understanding the relationship between the bacterium and atherosclerosis may now be started, with the hopes of developing preventative drugs or therapies.
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Creighton University
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Copyright is retained by the Author. A non-exclusive distribution right is granted to Creighton University and to ProQuest following the publishing model selected above.