Rapid assessment of MGMT promoter methylation status by droplet digital polymerase chain reaction on intraoperative smear preparation: A proof-of-concept study

Zachary Coty-Fattal; Heather Smith; Matthew McCord; Melissa Mejia-Bautista; David Dittman; Ditte Primdahl; Kyle Conway; Nicole Becker; Jared T Ahrendsen; Lucas Santana-Santos; Rudolph J Castellani; Madina Sukhanova; Lawrence J Jennings; Pouya Jamshidi

doi:10.1093/jnen/nlaf122

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Rapid assessment of MGMT promoter methylation status by droplet digital polymerase chain reaction on intraoperative smear preparation: A proof-of-concept study

Journal article

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Rapid assessment of MGMT promoter methylation status by droplet digital polymerase chain reaction on intraoperative smear preparation: A proof-of-concept study

Zachary Coty-Fattal, Heather Smith, Matthew McCord, Melissa Mejia-Bautista, David Dittman, Ditte Primdahl, Kyle Conway, Nicole Becker, Jared T Ahrendsen, Lucas Santana-Santos, …

Journal of neuropathology and experimental neurology, Vol.85(3), pp.280-284

03/01/2026

DOI: 10.1093/jnen/nlaf122

PMID: 41133796

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Abstract

MGMT promoter methylation is an important predictor of response to chemotherapy in adult-type diffuse gliomas. Currently, MGMT promoter methylation assessment requires formalin-fixed, paraffin-embedded (FFPE) tissue. We developed a droplet digital PCR (ddPCR) MGMT assay performed on bisulfite converted DNA utilizing primers targeting CpG positions with known clinical utility, which we validated against DNA methylation array (98.7% concordance). Smear preparations are routine for intraoperative neurosurgical consultation; thus, we sought to validate ddPCR using smear preparations. Thirty-six smears were collected during intraoperative consultation. For the first 28 samples, DNA from smears was extracted and bisulfite converted, and MGMT promoter methylation was quantified by ddPCR. These results were compared to DNA methylation array. For the final 8 samples, slides were H&E-stained and cover slipped for cellularity assessment prior to DNA extraction and directly compared with corresponding FFPE specimens. The smear ddPCR method showed concordance in 81.6% of cases, with a sensitivity of 85%, specificity of 100%, positive predictive value of 100% and negative predictive value of 81.3%. This novel approach will reduce turnaround time (approximately 2 days) over conventional MGMT analysis with FFPE and could provide clinically pertinent information for treatment planning and clinical trial enrollment options prior to discharge of the patient.

ddPCR

CNS tumor

molecular

droplet digital

Management

methylation

Details

Title: Subtitle: Rapid assessment of MGMT promoter methylation status by droplet digital polymerase chain reaction on intraoperative smear preparation: A proof-of-concept study
Creators: Zachary Coty-Fattal - Northwestern University
Heather Smith - Northwestern University
Matthew McCord - University of Virginia
Melissa Mejia-Bautista - Northwestern University
David Dittman - Northwestern University
Ditte Primdahl - Northwestern University
Kyle Conway - University of Michigan
Nicole Becker - University of Michigan
Jared T Ahrendsen - Northwestern University
Lucas Santana-Santos - Northwestern University
Rudolph J Castellani - Northwestern University
Madina Sukhanova - Northwestern University
Lawrence J Jennings - Northwestern University
Pouya Jamshidi - Northwestern University
Resource Type: Journal article
Publication Details: Journal of neuropathology and experimental neurology, Vol.85(3), pp.280-284
DOI: 10.1093/jnen/nlaf122
PMID: 41133796
NLM abbreviation: J Neuropathol Exp Neurol
ISSN: 0022-3069
eISSN: 1554-6578
Publisher: Oxford University Press
Language: English
Electronic publication date: 10/24/2025
Date published: 03/01/2026
Academic Unit: Pathology
Record Identifier: 9985016423702771

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