A qPCR technology for direct quantification of methylation in untreated DNA.

Autor: Bendixen KK; PentaBase A/S, Odense, Denmark. kkb@pentabase.com.; Epidemiology, Biostatistics and Biodemography, Department of Public Health, University of Southern Denmark, Odense, Denmark. kkb@pentabase.com., Mindegaard M; PentaBase A/S, Odense, Denmark., Epistolio S; Laboratory of Molecular Pathology, Institute of Pathology, Ente Ospedaliero Cantonale (EOC), Locarno, Switzerland., Dazio G; Laboratory of Molecular Pathology, Institute of Pathology, Ente Ospedaliero Cantonale (EOC), Locarno, Switzerland., Marchi F; Service of Neurosurgery, Neurocenter of the Southern Switzerland, Regional Hospital of Lugano, Lugano, Switzerland., Spina P; Laboratory of Molecular Pathology, Institute of Pathology, Ente Ospedaliero Cantonale (EOC), Locarno, Switzerland.; Department of Health Sciences, University of Eastern Piedmont, Novara, Italy., Arnspang EC; Department of Green Technology, University of Southern Denmark, Odense, Denmark., Soerensen M; Epidemiology, Biostatistics and Biodemography, Department of Public Health, University of Southern Denmark, Odense, Denmark., Christensen UB; PentaBase A/S, Odense, Denmark., Frattini M; Laboratory of Molecular Pathology, Institute of Pathology, Ente Ospedaliero Cantonale (EOC), Locarno, Switzerland., Petersen RK; PentaBase A/S, Odense, Denmark.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2023 Aug 24; Vol. 14 (1), pp. 5153. Date of Electronic Publication: 2023 Aug 24.
DOI: 10.1038/s41467-023-40873-y
Abstrakt: DNA methylation is important for gene expression and alterations in DNA methylation are involved in the development and progression of cancer and other major diseases. Analysis of DNA methylation patterns has until now been dependent on either a chemical or an enzymatic pre-treatment, which are both time consuming procedures and potentially biased due to incomplete treatment. We present a qPCR technology, EpiDirect®, that allows for direct PCR quantification of DNA methylations using untreated DNA. EpiDirect® is based on the ability of Intercalating Nucleic Acids (INA®) to differentiate between methylated and unmethylated cytosines in a special primer design. With this technology, we develop an assay to analyze the methylation status of a region of the MGMT promoter used in treatment selection and prognosis of glioblastoma patients. We compare the assay to two bisulfite-relying, methyl-specific PCR assays in a study involving 42 brain tumor FFPE samples, revealing high sensitivity, specificity, and the clinical utility of the method.
(© 2023. Springer Nature Limited.)
Databáze: MEDLINE
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