The 12 o’clock assay: an optimized dodecaplex droplet digital PCR assay for robust DNA methylation quantification and epigenetic clock-based age-predictions
DNA methylation quantification in age-prediction models relies on only a few techniques with inherent limitations and biases, reducing the accuracy and reproducibility. We developed a new droplet digital PCR assay format for accurate DNA methylation quantification, based on three LNA-containing hydrolysis probes to detect methylated and unmethylated alleles. Using a 12-plex assay we simultaneously quantified methylation at six CpGs (ASPA, C1orf132, CCDC102B, EDARADD, ELOVL2, FHL2) in 351 blood samples (age range: 0-95 years), showing strong correlations with age (|r|> 0.83). Several age-prediction models were developed, achieving high performance on testing sets (R<sup>2</sup>: 0.933-0.973, MAE: 3.18-5.18, RMSE: 4.71-7.27).
Institutions
- Conservatoire National des Arts et Métiers (FR)
- Institut de Radioprotection et de Sûreté Nucléaire (FR)
- Inserm (FR)
- Université Paris Cité (FR)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
- Fondation Jean Dausset-CEPH (FR)
- Sorbonne Université (FR)
- Centre National de Recherche en Génomique Humaine (FR)
- Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement (FR)
- Université Sorbonne Paris Nord (FR)
- Sorbonne Paris Cité (FR)
- Direction de la Recherche Fondamentale (FR)
- Génomique Bioinformatique et Applications (FR)
- French Clinical Research Infrastructure Network (FR)
- Centre de Recherche Épidémiologie et Statistique (FR)
- Institut Curie (FR)
Publication Details
- Journal
- Clinical Epigenetics
- Published
- 2026-03-24
- DOI
- https://doi.org/10.1186/s13148-026-02105-0
- Primary Topic
- Epigenetics and DNA Methylation
- Type
- article
- Field-Weighted Citation Impact
- 0.00