The application of pyrosequencing in periopathogen determination in aortic valves and in sulcular fluid
Abstract
Periodontitis may be a risk factor for inflammatory-degenerative diseases of heart and big vessels. The influence of periodontal pathogens in this respect needs further investigation.
Aims: The aim of this clinical-laboratory study was to test the presence of periodontal bacteria DNA in the samples of aortic valves extracted during their surgical replacement and to assess the concomitant presence of the same periodontal bacteria DNA in periodontal environment. Material and methods: Fifteen patients, scheduled for the aortic valve replacement surgery were involved in the study.
In the pre-operation stage they were periodontally examined and the samples of periodontal pockets contents were taken. Samples of impaired valves were taken during the surgical valve-replacement intervention.
It was very difficult to isolate microbial DNA from aortic valves even if the standard isolation method of microbial DNA from sulcular fluid samples yielded enough material.Therefore five different methods were used for the isolation of bacterial DNA from valves samples. Both samples were processed in molecular microbiologic techniques using the 454 pyrosequencing for the qualitative detection of selected periodontal bacteria.
The 454 pyrosequencing technique was used to detect the putative periodontal pathogens Tannerella forshytia, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, Eikenella corrodens, Treponema denticola and many others. Results: The 454 pyrosequencing technology showed a very infrequent presence of periopathogenic microbial DNA in aortic valves samples, on the other hand, samples from periodontal pockets contents of the same patients revealed the standard pattern of periopathogenic microflora.
Conclusions: The role of periopathogenic microflora in chronic inflammation process in aortic valves was not proved in this study.