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Evaluation of PCR primers to identify Porphyromonas endodontalis in apical periodontitis clinical samples
J Astorga1, M Hernández1,2, D Bravo3
1Laboratory of Periodontal Biology, Faculty of Dentistry, Universidad de Chile, Olivos 943, 8380492, Independencia, 8380000, Santiago, Chile.
Abstract:
We aimed to evaluate whether two commonly used PCR primers are effective to identify P. endodontalis and discriminate from other prevalent black-pigmented bacteria in apical periodontitis (AP). Endodontic canal samples from patients with asymptomatic AP (n = 20) were collected and cultured in anaerobiosis. Two primer sets to detect P. endodontalis were selected from the literature and first analyzed for their specificity in silico; and then tested on clinical isolates in vitro and finally, in apical exudates ex vivo. The identity of P. endodontalis was verified by PCR and Sanger sequencing with universal primers for bacterial V3-V6 regions 16S rDNA. Only one primer set showed specificity only for P. endodontalis clones in silico and also was specific for P. endodontalis in vitro and ex vivo.
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