title: Methylation-sensitive high-resolution melting in the context of legislative requirements for validation of analytical procedures for diagnostic applications.
abstract: Temperature gradient was first used to identify the methylation status of the DNA sequence over 10 years ago; however, the initially published protocol was shown to have poor analytical sensitivity. Recent developments in the field of DNA melting technologies, combined with the identification of procedures to overcome the sensitivity issues in the PCR-based methylation detection applications, led to the development of the methylation-sensitive high-resolution melting (MS-HRM) protocol. This protocol allows for highly sensitive detection of methylation levels in a labor- and cost-efficient fashion. Moreover, it enables investigation of methylation status of imprinted loci as well as identification of heterogeneous methylation. The MS-HRM technology is being increasingly applied in research laboratories and has a potential for future application in diagnostic settings. The focus of this article is to describe the development of the HRM technology for methylation analyses and evaluate the diagnostic applicability of the MS-HRM technology.
authors: Lundbeck Foundation Centre for International Research in Radiation Oncology, Aarhus C, Denmark. wojdacz@humgen.au.dk
authors: Tomasz K
authors: TK
authors: Wojdacz
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics; D004247:DNA / Q000737:chemistry*; D019175:DNA Methylation; D057069:Diagnostic Test Approval / Q000331:legislation & jurisprudence / Q000592:standards; D006801:Humans; D057230:Limit of Detection; D025202:Molecular Diagnostic Techniques / Q000592:standards*; D009369:Neoplasms / Q000175:diagnosis; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; D004247:DNA