title: Clinical Neuropathology practice news 1-2014: pyrosequencing meets clinical and analytical performance criteria for routine testing of MGMT promoter methylation status in glioblastoma.
abstract: Testing of the MGMT promoter methylation status in glioblastoma is relevant for clinical decision making and research applications. Two recent and independent phase III therapy trials confirmed a prognostic and predictive value of the MGMT promoter methylation status in elderly glioblastoma patients. Several methods for MGMT promoter methylation testing have been proposed, but seem to be of limited test reliability. Therefore, and also due to feasibility reasons, translation of MGMT methylation testing into routine use has been protracted so far. Pyrosequencing after prior DNA bisulfite modification has emerged as a reliable, accurate, fast and easy-to-use method for MGMT promoter methylation testing in tumor tissues (including formalin fixed and paraffin-embedded samples). We performed an intra- and inter-laboratory ring trial which demonstrates a high analytical performance of this technique. Thus, pyrosequencing- based assessment of MGMT promoter methylation status in glioblastoma meets the criteria of high analytical test performance and can be recommended for clinical application, provided that strict quality control is performed. Our article summarizes clinical indications, practical instructions and open issues for MGMT promoter methylation testing in glioblastoma using pyrosequencing.
authors.affiliation: Department of Medicine I, Medical University of Vienna, Vienna, Division of Pathology, Medical University of Innsbruck, Innsbruck, Comprehensive Cancer Center - CNS Tumors Unit (CCC-CNS), Medical University of Vienna, Vienna, Molecular Diagnostics, Austrian Institute of Technology, Department of Radiotherapy, Department of Neurosurgery, and Institute of Neurology, Medical University of Vienna, Vienna, Austria.
authors.forename: Matthias
authors.initials: M
authors.lastname: Preusser
authors.forename: Anna S
authors.initials: AS
authors.lastname: Berghoff
authors.forename: Claudia
authors.initials: C
authors.lastname: Manzl
authors.forename: Martin
authors.initials: M
authors.lastname: Filipits
authors.forename: Andreas
authors.initials: A
authors.lastname: Weinhäusel
authors.forename: Walter
authors.initials: W
authors.lastname: Pulverer
authors.forename: Karin
authors.initials: K
authors.lastname: Dieckmann
authors.forename: Georg
authors.initials: G
authors.lastname: Widhalm
authors.forename: Adelheid
authors.initials: A
authors.lastname: Wöhrer
authors.forename: Engelbert
authors.initials: E
authors.lastname: Knosp
authors.forename: Christine
authors.initials: C
authors.lastname: Marosi
authors.forename: Johannes A
authors.initials: JA
authors.lastname: Hainfellner
mesh_terms: D000328:Adult; D000368:Aged; D001932:Brain Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000378:metabolism*; D045643:DNA Repair Enzymes / Q000378:metabolism*; D011756:Diphosphates; D005909:Glioblastoma / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008163:Luminescent Measurements; D021141:Nucleic Acid Amplification Techniques; D011237:Predictive Value of Tests; D011401:Promoter Regions, Genetic; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
chemical_list: D011756:Diphosphates; D013447:Sulfites; D025521:Tumor Suppressor Proteins; C107241:diphosphoric acid; D015254:DNA Modification Methylases; C515491:MGMT protein, human; D045643:DNA Repair Enzymes; C042345:hydrogen sulfite