20.500.12556/DiRROS-28991 Digital PCR outperforms quantitative real-time PCR for the detection and quantification of major periodontal pathobionts Background: This study comparatively evaluated the analytical and diagnostic performance of a multiplex digital polymerase-chain reaction (dPCR) assay and a quantitative real-time PCR (qPCR) for the simultaneous detection and quantification of periodontal pathobionts: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum. Materials and methods: Subgingival plaque samples from 20 periodontitis patients and 20 periodontally healthy controls were analyzed. Several analytical parameters of the dPCR assay, optimized using DNA standards, were assessed versus qPCR: dynamic range linearity, precision, accuracy, prevalence, sensitivity, specificity, and concordance. The statistical analyses included Mann-Whitney U test, Wilcoxon test, McNemar's test, and Bland-Altman plots. Results: dPCR showed high linearity (R2 > 0.99) and lower intra-assay variability (median CV%: 4.5%) than qPCR (p = 0.020), with comparable accuracy and agreement. dPCR demonstrated superior sensitivity, detecting lower bacterial loads, particularly for P. gingivalis and A. actinomycetemcomitans. The Bland-Altman plots highlighted good agreement at medium/high loads but discrepancies at low concentrations (< 3 log10Geq/mL), resulting in qPCR false negatives and a 5-fold underestimation of the prevalence of A. actinomycetemcomitans in periodontitis patients. High concordance between the assays was observed for F. nucleatum across both study groups. Conclusions: dPCR outperformed qPCR for quantifying periodontal pathobionts and had superior sensitivity and precision, making it particularly effective in detecting low-level bacterial loads. digital PCR oral microbiology periodontal disease quantitative real-time PCR subgingival plaque digitalni PCR oralna mikrobiologija parodontalna bolezen kvantitativna PCR v realnem času subgingivalni plak true false true Angleški jezik Slovenski jezik Neznano 2026-04-15 14:56:33 2026-04-15 14:56:34 2026-04-16 04:06:23 0000-00-00 00:00:00 2025 0 0 Nasl. z nasl. zaslona; Opis vira z dne: 7. 8. 2025; str. 1-13 iss. 1, [article no.] 2537439 Vol. 17 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616.31 2000-2297 10.1080/20002297.2025.2537439 245171203 RAZ_Munjakovic_Haris_2025.pdf RAZ_Munjakovic_Haris_2025.pdf 1 AF3DA315155973612B33B71EEBC82778 0b408b4ae846b4f609b47f1b5d75544318416382c3e7a46df4d71153dbb82ad4 a6973cc5-38ca-11f1-9cc3-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=43272 https://www.tandfonline.com/doi/10.1080/20002297.2025.2537439?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmed 1 8922fdc3-38ca-11f1-9cc3-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=43271 Univerzitetni klinični center Ljubljana 0 0 0