Direct bisulphite conversion of cervical samples for DNA methylation analysis
| Autor: | Saskia Doorn, Lisanne Verhoef, Kate Cuschieri, Arno Floore, Daniëlle A M Heideman, Ramya Bhatia, Renske D.M. Steenbergen, Chris J.L.M. Meijer, Albertus T. Hesselink |
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| Přispěvatelé: | Pathology, CCA - Cancer biology and immunology, CCA - Imaging and biomarkers |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Cancer Research
Sodium bisulphite Uterine Cervical Neoplasms Biology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Methylation analysis Humans Sulfites 030212 general & internal medicine Molecular Biology Gene Papillomavirus Infections Methylation DNA DNA Methylation Molecular biology genomic DNA chemistry 030220 oncology & carcinogenesis Liquid-based cytology DNA methylation Cytokines Female |
| Zdroj: | Epigenetics. Landes Bioscience Verhoef, L, Floore, A, Doorn, S, Cuschieri, K, Bhatia, R, Hesselink, B, Meijer, C J L M, Steenbergen, R D M & Heideman, D A M 2021, ' Direct bisulphite conversion of cervical samples for DNA methylation analysis ', Epigenetics . https://doi.org/10.1080/15592294.2021.1992911 |
| ISSN: | 1559-2294 |
| Popis: | Sodium bisulphite conversion of DNA to separate methylated from unmethylated cytosines is a standard for methylation analysis. This study evaluated a direct cell conversion protocol on cervical samples as alternative to isolated genomic DNA as input. Clinician-collected cervical samples (n = 120) were subjected to a direct conversion protocol, or genomic DNA was isolated with a fixed amount used for subsequent bisulphite conversion. Converted samples were compared for ACTB control gene and methylation of FAM19A4 and miR124-2 genes using quantitative methylation-specific PCR (QIAsure Methylation Test). Direct conversion resulted in a high success rate, i.e., 119/120 (99.2%) samples reported a valid test result. ΔΔCq values of FAM19A4 and miR124-2 were significantly correlated between both protocols (Spearman Rho 0.708 and 0.763, respectively, all p-values = 0.000). Agreement between both the bisulphite protocols was demonstrated by Bland–Altman plots. A direct cell conversion protocol shows good technical and analytical performance and offers a streamlined workflow for methylation analysis. |
| Databáze: | OpenAIRE |
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