Prospects application of polypyrrole-based immunosensor to Porphyromonas gingivalis quantification in subgingival...

Periodontitis still poses a serious threat to oral and systemic health condition of humans. The proportion of the main pathogenic bacteria change in localized sites was associated with the initiation of the disease process. However, the limitations of microbiological diagnostic aids rendered the diagnosis of active periodontitis status point-of-care or chair-side based on microbiological data difficult.

Porphyromonas gingivalis, a major putative etiological agent in the initiation and progression of chronic periodontitis, was used as the experimental subject. An immunosensor based on polypyrrole-coated interdigitated array microelectrodes was developed to quantify Porphyromonas gingivalis in pure culture, gingival crevicular fluid and saliva samples. The regression equation for the normalized impedance change (NIC) versus Porphyromonas gingivalis concentration (C) was measured. The correlation between results of the immunosensor and quantitative real-time PCR method in quantifying Porphyromonas gingivalis in subgingival plaque samples was evaluated.

Results of the study revealed that the lowest detection limits of the immunosensor was 1.9 x 10(4), 2.7 x 10(5), and 2.7 x 10(6) cells/mL in pure culture, gingival crevicular fluid, and saliva samples respectively. The values determined using the immunosensor strongly correlated with those obtained using quantitative real-time PCR method (R2 = 0.91, p < 0.05). The immunosensor did not require any labels and amplification steps, and the total detection time from sampling to measurement was less than one hour.

The immunosensor developed in the present study offered some insight into monitoring the change in the number of periodontal bacteria chair-side during routine clinical practice.