Quantification by qPCR of Pathobionts in Chronic Periodontitis: Development of Predictive Models of Disease Severity at Site-Specific Level

Abstract

Currently, there is little evidence available on the development of predictive models for the diagnosis or prognosis of chronic periodontitis based on the qPCR quantification of subgingival pathobionts. Our objectives were to: (1) analyze and internally validate pathobiont-based models that could be used to distinguish different periodontal conditions at site-specific level within the same patient with chronic periodontitis; (2) develop nomograms derived from predictive models. Subgingival plaque samples were obtained from control and periodontal sites (probing pocket depth and clinical attachment loss <4 mm and >4 mm, respectively) from 40 patients with moderate-severe generalized chronic periodontitis. The samples were analyzed by qPCR using TaqMan probes and specific primers to determine the concentrations of <i>Actinobacillus actinomycetemcomitans (Aa)</i>, <i>Fusobacterium nucleatum (Fn)</i>, <i>Parvimonas micra (Pm)</i>, <i>Porphyromonas gingivalis (Pg)</i>, <i>Prevotella intermedia (Pi)</i>, <i>Tannerella forsythia (Tf)</i>, and <i>Treponema denticola (Td)</i>. The pathobiont-based models were obtained using multivariate binary logistic regression. The best models were selected according to specified criteria. The discrimination was assessed using receiver operating characteristic curves and numerous classification measures were thus obtained. The nomograms were built based on the best predictive models. Eight bacterial cluster-based models showed an area under the curve (AUC) ≥0.760 and a sensitivity and specificity ≥75.0%. The <i>PiTfFn</i> cluster showed an AUC of 0.773 (sensitivity and specificity = 75.0%). When <i>Pm</i> and <i>AaPm</i> were incorporated in the <i>TdPiTfFn</i> cluster, we detected the two best predictive models with an AUC of 0.788 and 0.789, respectively (sensitivity and specificity = 77.5%). The <i>TdPiTfAa</i> cluster had an AUC of 0.785 (sensitivity and specificity = 75.0%). When <i>Pm</i> was incorporated in this cluster, a new predictive model appeared with better AUC and specificity values (0.787 and 80.0%, respectively). Distinct clusters formed by species with different etiopathogenic role (belonging to different Socransky's complexes) had a good predictive accuracy for distinguishing a site with periodontal destruction in a periodontal patient. The predictive clusters with the lowest number of bacteria were <i>PiTfFn</i> and <i>TdPiTfAa</i>, while <i>TdPiTfAaFnPm</i> had the highest number. In all the developed nomograms, high concentrations of these clusters were associated with an increased probability of having a periodontal site in a patient with chronic periodontitis.