Real-time methylation-specific polymerase chain reaction for MGMT promoter methylation clinical testing in glioblastoma an alternative detection method for a heterogeneous process

Cristiane M. Ida, Malinda L. Butz, Robert B. Jenkins, Jann N. Sarkaria, Gaspar J. Kitange, Caterina Giannini, Benjamin R. Kipp

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Objectives: To develop and evaluate a real-time methylation-specific polymerase chain reaction (RT-MSP) MGMT assay, with a particular focus on small biopsies and indeterminate testing results. Methods: We assessed formalin-fixed paraffin-embedded glioblastoma or gliosarcoma specimens (n = 641). A testvalidation group (n = 51) with previously obtained reference laboratory (RL) results was used to determine performance characteristics of the RT-MSP assay. An indeterminate (equivocal) category was established for cases that could not be clearly classified as positive or negative. Results: Overall agreement of RT-MSP and RL results was 91% (41/45 nonindeterminate cases). Discordant cases were tested by pyrosequencing, and results were most concordant with RT-MSP. Among cases with limited amounts of tissue (n = 7), six yielded valid results by RT-MSP (all negative); the single invalid result consisted of a stereotactic biopsy specimen obtained 14 years prior. A subset of indeterminate cases obtained during clinical testing (n = 18/575 [3%]) was also evaluated by pyrosequencing and showed a heterogeneous pattern of methylation across the eight interrogated CpG sites. Conclusions: The RT-MSP assay that we developed in-house is a robust clinical detection method for the heterogeneous process of MGMT promoter methylation in glioblastoma.

Original languageEnglish (US)
Pages (from-to)296-307
Number of pages12
JournalAmerican journal of clinical pathology
Volume148
Issue number4
DOIs
StatePublished - Oct 1 2017

Keywords

  • Astrocytoma
  • Brain
  • Epigenetics
  • Glioma
  • Molecular

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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