Detection of putative periodontal pathogens (PPP's) by PCR is usually determined with samples of subgingival plaque collected using a curette or paper-point. Due to the invasiveness of these collection methods the services of a dentist is usually required. Objective: To investigate the use of a non-invasive oral lavage sampling method to detect 4 PPP's. Methods: Oral lavage fluid was obtained from 8 periodontal maintenance patients by having them swish vigorously with 5 ml of sterile water for 10 seconds then expectorate into a sterile container. This procedure was repeated twice to give three lavage samples. The samples were placed on ice and within 1 hour they were centrifuged at 10,000 x g for 10 minutes in order to pellet cells and particulates. The supernatants were removed and the pellets suspended in 200 ml of a lysis buffer and heated at 55°C for 2 hours, then boiled for 10 minutes, cooled on ice then centrifuged at 16,000 x g for 5 minutes. The DNA in 1 ml of the supernatant was used as template for a nested PCR procedure designed to detect Treponema denticola (TD), Bacteroides forsythus (BF), Porphyromonas gingivalis (PG), or Actinobacillus actinomycetemcomitans (AA). Results: All subjects were positive for at least 1 species; no subject was positive for all 4 species; 4 subjects were positive for two species; and 3 subjects were positive for 3 species. The carriage rates for each species were 6/8 for TD and BF and 3/8 for AA and PG. All of these patients harbored at least one species of the “Red Complex” regardless of periodontal pocket probing depths. Conclusions: This non-invasive sampling method was successful for the detection of PPP's by PCR. The method will allow unskilled personnel to collect samples from multiple subjects to screen for the carriage of PPP's by PCR.

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