Detection of Black-Pigmented Bacteria in Pus of Acute Endodontic Infection by Multiplex Polymerase Chain Reaction
The aim of this pilot study was to determine whether a multiplex polymerase chain reaction (PCR) protocol could detect DNA from four different pathogenic bacteria, including Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia and Prevotella nigrescens in 10 pus exudate samples from root canals. The conventional PCR together with a pair of universal primers that was specific for 16s ribosomal RNA was initially conducted to show the presence of any bacterial DNA in all specimens. Then, the primers for multiplex PCR were specifically designed to amplify the DNA of two different bacterial species at a time, i.e. Porphyromonas endodontalis and Porphyromonas gingivalis versus Prevotella intermedia and Prevotella nigrescens. The results showed that two of the specimens (20%) failed to show the presence of DNA in any of the four bacteria. The percentage of sample where bacterial DNA was detected for three, two or one species was 40, 10 or 30%, respectively. Moreover, the presence of DNA was detected in Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella nigrescens and Prevotella intermedia by 70, 40, 40 and 20% of the sample, respectively. In summary, the present pilot study shows the effectiveness of multiplex PCR for detecting distinct bacterial DNA in endodontic specimens. However, additional studies with a larger number of samples are still required to corroborate these findings.
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