Detection of Periodontal Bacteria in Atheromatous Plaques by Nested Polymerase Chain Reaction

This research was originally published in 2011 by Elena Figuero , María Sánchez-Beltrán, Susana Cuesta-Frechoso, Jose María Tejerina, Jose Antonio del Castro, Jose María Gutiérrez, David Herrera, Mariano Sanz.

We have curated this article as a reference point for The Larkin Protocol.


Background. In recent years, increasing evidence regarding the potential association between periodontal diseases and cardiovascular diseases has been elicited. The available evidence underlines the importance of detecting periodontal pathogens on atheromatous plaques as the first step in demonstrating the causal relationship between both entities. It is the main aim of this investigation to detect periodontitis-associated bacteria from carotid artery atheromatous plaques recovered from patients who received an endarterectomy, using strict sample procurement and laboratory procedures.

Methods. Atheromatous plaques from endarterectomies from carotid arteries were scraped, homogenized and bacterial DNA was extracted. In order to obtain a representative concentration of amplicons, two amplifications of the bacterial 16S ribosomal-RNA gen were carried out for each sample with universal eubacteria primers by polymerase chain reaction (PCR). Nested-PCR with specific primers for the target bacteria was performed next. Statistical tests included χ2 test. Results. Forty-two atheromatous plaques were analyzed. All of them were positive for at least one target bacterial species. The bacterial species most commonly found in atheromatous plaques was Porphyromonas gingivalis (78.57%, 33/42), followed by Aggregatibacter actinomycetemcomitans (66.67%, 28/42), Tannerella forsythia (61.90%, 26/42), Eikenella corrodens (54.76%, 23/42), Fusobacterium nucleatum (50.00%, 21/42) and Campylobacter rectus (9.52%, 4/42). The simultaneous presence of various bacterial species within the same specimen was a common observation.

Conclusion. Within the limitations of this study, the presence of DNA from periodontitisassociated bacteria in carotid artery atheromatous plaques retrieved by endarterectomy was confirmed.

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