Comparison of template preparation methods for rapid detection of Porphyromonas gingivalis

Article Type: Full Length Research Article Polymerase chain reaction (PCR) and loop mediated isothermal amplification (LAMP) represent a major advance in diagnostic methods in terms of speed and sensitivity. However, standard methods for sample preparation prior to PCR and LAMP are laborious and time consuming. Three template preparation methods (crude template, boiling, and commercial DNA extraction kit) were compared for detection of Porphyromonas gingivalis using PCR and LAMP assays. A total of eight sub-gingival plaques were evaluated for the presence of P. gingivalis. Both PCR and LAMP reactions revealed the presence of P. gingivalis in the plaque samples. When tested by PCR, 7/8 (87%) of samples extracted by a simple boiling method were positive, whereas 6/8 (75%) were positive after extraction, using commercial kit and 5/8 (62%) were positive after extraction, using a direct method. Whereas when tested by LAMP, all eight (100%) samples were positive after extraction by boiling and direct methods and seven (87%) were positive using a commercial kit. Higher sensitivity (100%) was observed for LAMP assay compared to those from PCR assay. In conclusion, crude supernatant of subgingival plaque appears to represent a rapid, simple and cheaper method when LAMP assay was used for P. gingivalis detection. ©2016 BluePen Journals Ltd. All rights reserved