EpiDirect® MGMT Methylation Assay a direct qPCR, Bisulfite-Free.
EpiDirect® MGMT Methylation qPCR Assay, CE IVD, IVDR
The EpiDirect® MGMT Methylation qPCR Assay represents a new and innovative approach allowing for direct quantification of MGMT promoter methylation by qPCR without prior DNA conversion.
The EpiDirect® technology is based on our unique INA® chemistry and includes an EpiPrimer™ oligonucleotide that specifically binds and primes amplification of methylated target DNA.
The assay is able to quantify MGMT promoter methylation using standard real-time PCR instruments* in less than two hours with as little as 0.5 ng of unconverted input DNA.
The assay is compatible with FFPE and fresh-frozen glioblastoma samples and eliminates the need for time-consuming bisulfite conversion.
- Direct DNA methylation analysis by qPCR without DNA pretreatment
- High sensitivity: detects methylation levels as low as 1.5%
- Standard qPCR instrument procedure*
- Results in less then 2 hours
- Minimal hands-on time
- 0.5-50 ng input DNA
- CE IVD
- IVDR certified^
*Validated on CFX96/CFX Opus 96 (Bio-Rad), QuantStudio™ 5 (ABI) and BaseTyper™ 48.4 (PentaBase) Real-Time PCR Systems
^Conformity assessment by Notified Body Number 2797
Single Registration Number: DK-MF-000023157
Product Details
Reference Number:
8300
Product Name:
EpiDirect® MGMT Methylation qPCR Assay
Product Description:
Includes PCR master mix and oligonucleotide mix for quantification of MGMT promoter methylation, Positive Control, Negative Control and Standard Curve Controls
Format:
Ready-to-Use
Reactions:
48
Intended Use
EpiDirect® MGMT Methylation qPCR Assay is a real-time polymerase chain reaction (PCR) assay intended to detect and quantify the methylation status of the human O6-methylguanine-DNA methyltransferase (MGMT) gene promoter. The assay is intended for use with purified human genomic DNA extracted from formalin-fixed paraffin-embedded brain tumor samples that, without further pre-treatment, can be analysed directly by semi-automatic real-time PCR systems. The assay should be used quantitatively by applying the included standard curve. The methylation status of the MGMT promoter determined by the EpiDirect® MGMT Methylation qPCR assay can be used for stratification of brain tumor patients in accordance with current clinical guidelines.
The assay is intended for use by healthcare professionals or qualified laboratory personnel specifically instructed and trained in the techniques of real-time PCR as well as proficient in handling biological samples.
Product Specifications
Procedure:
Real-time PCR-based
Validated Real-Time PCR instruments:
BaseTyper™ 48.4 Quiet HRM (PentaBase Ref. No. 754), CFX96 and CFX Opus 96 (Bio-Rad) and QuantStudio™ 5 Real-Time PCR Systems
PentaBase technologies:
EpiPrimer™, HydrolEasy® Probe, and SuPrimer™
Genomic target:
DMR2 region CpG sites 75-78 in exon 1 of the MGMT promoter
Sample types:
Formalin-fixed paraffin-embedded (FFPE) or fresh-frozen solid brain tumor biopsies
The sample tumor cell percentage:
At least 20%
Sample DNA extraction methods:
Relevant commercially avaliable solid biospy/FFPE DNA extraction kits
Sample input:
0.1-10 ng/µL of purified DNA directly from the sample that has not been otherwise chemically manipulated
PCR run time:
Less than 2 hours
Instructions For Use
For the latest version of the Instructions For Use (IFU) – Click here –
Publications
A qPCR technology for direct quantification of methylation in untreated DNA
Bendixen et al. – Nature Communications – 2023
Related files
A Revolutionary Approach to DNA Methylation Testing
Current methods for evaluating DNA methylation typically rely on sodium bisulfite or other chemical and enzymatic conversion of cytosines. However, variability in conversion efficiency and DNA degradation caused by chemical treatment can lead to false or inconclusive results. In particular, bisulfite treatments often cause biased degradation of unmethylated DNA, which may overestimate methylation levels.
In contrast, the EpiDirect® technology allows for direct quantification of DNA methylation by PCR without any prior chemical or enzymatic treatment. As a result, it eliminates artefacts related to incomplete conversion of cytosines and DNA degradation, while vastly reducing hands-on time and total time to result.
As a result, EpiDirect® MGMT provides accurate, reproducible, and efficient methylation analysis, making it ideal for both research and clinical applications.
How The EpiDirect® Technology Works
The technology relies on a unique DNA analogue chemistry called intercalating pseudo-nucleotides (IPN). These special oligonucleotides, also known as intercalating nucleic acids (INAs®), bind more strongly and specifically to target DNA than standard oligonucleotides.
Central to the system is the EpiPrimer™, which is designed to selectively amplify methylated DNA. Each EpiPrimer™ has three key parts:
- Anchor sequence: Binds to the methylated DNA region.
- Starter sequence: Initiates replication of the methylated target.
- Loop sequence: Supports priming of the remaining DNA during PCR cycles.
Together, the anchor and starter sequences drive initial replication, while the loop and starter sequences handle the later PCR cycles. As a result, the assay achieves accurate and efficient amplification without chemical treatments.
Moreover, this approach reduces errors, saves time, and is easier to use than traditional bisulfite-based methods. In addition, it works on standard PCR instruments, making it practical for both research and clinical labs.
