Real-Time PCR Quantification of Periodontal Pathogens in Diabetic Kuwaiti Children
Objectives: Prevalence of type-1 diabetes in children is increasing worldwide. A link has appeared between diabetes and periodontal diseases. Although diabetes in Kuwaiti children is common, no information is available of their periodontal microbiota. The aiom of this study was to assess the levels of major periodontal pathogens in diabetic and non-diabetic Kuwaiti children with gingivitis.
Methods: 6 children (14-18 years old) were included; 3 diabetics (HbA1C>7%) and 3 controls (HbA1C≤5.5%). None had received antibiotics during the preceding 3 months. 8 subgingival paper point samples were collected per child. Bacterial DNA was purified using DNeasy kit (Qiagen) and quantified on ABI Fast 7900 HT RT-PCR machine using Power SYBR Green” kit and species-specific primers. The detection limit for all target species was 102 CFU/ml.
Results: The detection rates among diabetics versus controls were for Aggregatibacter actinomycetemcomitans (Aa; 2 vs. 3), Porphyromonas gingivalis (Pg; 2 vs. 3), Prevotella intermedia (Pi; 2 vs. 1), Prevotella nigrescens (Pn; 2 vs. 2), Fusobacterium nucleatum (Fn; 3 vs. 3), Campylobacter rectus (Cr; 3 vs. 3), and Parvimonas micra (Pm; 2 vs. 2). Pg, Fn and Cr quantities were low (≤103 CFU/ml) in all children and did not significantly (P>0.05) differ between the groups. Aa and Pm neither differed (P>0.05) between the groups, although their mean quantities were higher in children with [(Aa: 1.15x105 (1.9x105), Pm: 2.4x104 (3.4x104) CFU/ml] than without [(Aa: 1.6x104 (7.3x103), Pm: 1x104 (9x103) CFU/ml] diabetes. Mean (SD) quantities of Pi and Pn were elevated [5x105 (7x105) and 5.6x105 (7x105) CFU/ml] in 2 of 3 patients in the diabetes group, but the difference from the non-diabetes group did not reach statistical significance. The diabetic with the lowest number of target bacteria also had the lowest HbA1C.
Conclusions: While the target species seem common, diabetics may support peak quantities.