| Title: | Digital PCR outperforms quantitative real-time PCR for the detection and quantification of major periodontal pathobionts |
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| Authors: | ID Munjaković, Haris (Author)
ID Povšič, Katja (Author)
ID Poljak, Mario (Author)
ID Seme, Katja (Author)
ID Gašperšič, Rok (Author)
ID Skubic, Lucijan (Author) |
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| Files: |  PDF - Presentation file, download (3,83 MB)
MD5: AF3DA315155973612B33B71EEBC82778
 URL - Source URL, visit 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
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| Language: | English |
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| Typology: | 1.01 - Original Scientific Article |
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| Organization: |  UKC LJ - Ljubljana University Medical Centre
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| Abstract: | 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. |
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| Keywords: | digital PCR, oral microbiology, periodontal disease, quantitative real-time PCR, subgingival plaque |
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| Publication status: | Published |
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| Publication version: | Version of Record |
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| Year of publishing: | 2025 |
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| Number of pages: | str. 1-13 |
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| Numbering: | Vol. 17, iss. 1, [article no.] 2537439 |
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| PID: | 20.500.12556/DiRROS-28991  |
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| UDC: | 616.31 |
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| ISSN on article: | 2000-2297 |
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| DOI: | 10.1080/20002297.2025.2537439 |
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| COBISS.SI-ID: | 245171203 |
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| Note: | Nasl. z nasl. zaslona;
Opis vira z dne: 7. 8. 2025;
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| Publication date in DiRROS: | 15.04.2026 |
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| Views: | 53 |
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| Downloads: | 19 |
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