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ME012 MGMT-IDH-TERT

SALSA MLPA Probemix ME012 MGMT-IDH-TERT detects copy number variations and methylation status of the MGMT gene, as well as selected SNVs.

Specifications

Contents: 31 MLPA probes, including 8 probes for MGMT, which all provide information on the methylation status, and probes for IDH1 (p.R132H and p.R132C), IDH2 (p.R172K and p.R172M), and TERT (C228T and C250T).

Tissue: human genomic DNA, including DNA from FFPE tissue.

Application: research on gliomas.

For research use only (RUO). Not for use in diagnostics.

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Key related products

P088 Oligodendroglioma 1p-19q

Contains probes for copy number detection of 1p and 19q chromosomal regions and for IDH1 (p.R132C, p.R132H) and IDH2 (p.R172K, p.R172M) point mutation detection.

P105 Glioma

Contains probes for PDGFRA, EGFR, CDKN2A, PTEN, TP53, CDK4-MIR26A2- MDM2 and NFKBIA genes.

P370 BRAF-IDH1-IDH2

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General information

The SALSA MLPA Probemix ME012 MGMT-IDH-TERT is a research use only (RUO) assay for the detection of aberrant methylation of one or more sequences of the MGMT gene, as well as deletions/gains in the aforementioned region. This probemix can also be used to detect the presence of IDH1 (p.R132H=c.395G>A and p.R132C=c.394C>T), IDH2 (p.R172K=c.515G>A and p.R172M=c.515G>T) and TERT promoter (C228T and C250T) point mutations in a DNA sample.

Gliomas, glioneuronal and neuronal tumours are the most common central nervous system (CNS) neoplasms, subdivided into six groups according to the fifth edition of the WHO classification of CNS tumours: adult-type diffuse gliomas (encompassing oligodendroglioma, glioblastoma and astrocytoma), paediatric-type diffuse low-grade gliomas, paediatric-type diffuse high-grade gliomas, circumscribed astrocytic gliomas, glioneuronal and neuronal tumours, ependymomas (Torp et al. 2022; Gritsch et al. 2022). Molecular genetic characteristics play an increasingly major role in classification, diagnostics and prognosis of CNS tumours.

IDH1 and IDH2 mutation (IDH-mutation) status represents an important diagnostic and prognostic marker in gliomas (Riemenschneider et al. 2010, van den Bent et al. 2010). The presence of IDH-mutation is suggested to associate with favourable prognosis and a longer survival of glioma patients (Sanson et al. 2009, Zou et al. 2013). The IDH1/2 mutations described are not activating or inactivating, but probably result in altered enzymatic properties (Hartmann et al. 2009). IDH-mutation is a marker for glioma classification since 2016, defining glioblastomas as IDH-mutant or IDH-wildtype (Wesseling and Capper 2018).

Point mutations in the TERT promoter region generate novel transcription factor binding sites and thus increase the expression of the telomerase enzyme encoded by TERT. Common TERT promoter mutations are known as C228T and C250T, referring to C>T transitions at hg19/GCRh37 chr5:1295228 and chr5:1295250 positions, respectively. These mutations are predominantly present in oligodendroglioma and are associated with poor prognosis and reduced survival in the absence of IDH-mutation (Labussière et al. 2014). TERT promoter mutation, in combination with IDH-mutation and 1p/19q codeletion, is characteristic of oligodendroglioma. Absence of TERT promoter mutation, coupled with the presence of IDH-mutation, designates astrocytoma (Cahill et al. 2015; Eckel-Passow et al. 2015).

CpG-islands are located in or near the promoter region or other regulatory regions of approximately 50% of human genes. Aberrant methylation of CpG-islands has been shown to be associated with transcriptional inactivation of genes in a wide spectrum of human cancers. These genes are frequently silenced by methylation in tumours, but are unmethylated in blood-derived DNA. In addition, DNA methylation analysis can indicate in some cases from which type of tissue the tumour was derived.

Hypermethylation in the promoter region of the MGMT gene, encoding for the DNA repair enzyme O6-methylguanine DNA methyltransferase, is an important prognostic marker and predictor for response to treatment in glioma with alkylating agents such as temozolomide (Weller et al. 2009, Hegi et al. 2005, Pegg 1990). Assessment of both the IDH-mutation and the MGMT methylation status is proposed to be used as a combined predictor for glioblastoma patient survival (Wick et al. 2013). Combined assessment of IDH-mutation and MGMT methylation status is suggested to predict survival in glioblastoma better than either biomarker alone (Molenaar et al. 2014). The presence of TERT promoter mutation in combination with unmethylated MGMT defines glioblastoma with the poorest prognosis (Arita et al. 2016).

More information is available at https://www.ncbi.nlm.nih.gov/books/NBK558954/ and WHO Classification of Central Nervous System Tumours, 5th Edition, Volume 6.

Regulatory status

SALSA MLPA Probemix ME012 MGMT-IDH-TERT is for research use only (RUO) in all territories.

SALSA Sample DNA for this product

SALSA Binning DNA SD094 is an artificial DNA sample with a signal for all probes in the ME012 MGMT-IDH-TERT probemix. Inclusion of a reaction with SD094 in initial experiments and in experiments following a change in electrophoresis conditions is recommended to aid in the creation of a bin set that links peaks to the probes that produce them. Binning DNA cannot be used as a reference sample in the MLPA data analysis, and cannot be used to quantify the signals of mutation-specific probes.

A vial of SALSA Binning DNA SD094 is included with every order of the ME012 MGMT-IDH-TERT probemix, but it is possible to order additional vials separately.

For more information, see the product description.

Product documentation

Version B1

Other versions

Contact us for earlier versions.

List prices

Product

Item no.
Description
Technology
Price
ME012-025R
SALSA MLPA Probemix ME012 MGMT-IDH-TERT – 25 rxn
MS-MLPA
€ 296.00
ME012-050R
SALSA MLPA Probemix ME012 MGMT-IDH-TERT – 50 rxn
MS-MLPA
€ 580.00
ME012-100R
SALSA MLPA Probemix ME012 MGMT-IDH-TERT – 100 rxn
MS-MLPA
€ 1135.00

Required reagents

A general SALSA MLPA Reagent Kit and SALSA HhaI are required for MS-MLPA experiments (to be ordered separately).

Item no.
Description
Technology
Price
EK1-FAM
SALSA MLPA Reagent Kit – 100 rxn – FAM (6 vials)
MLPA MS-MLPA
€ 361.00
EK1-CY5
SALSA MLPA Reagent Kit – 100 rxn – Cy5 (6 vials)
MLPA MS-MLPA
€ 361.00
EK5-FAM
SALSA MLPA Reagent Kit – 500 rxn – FAM (5×6 vials)
MLPA MS-MLPA
€ 1658.00
EK5-CY5
SALSA MLPA Reagent Kit – 500 rxn – Cy5 (5×6 vials)
MLPA MS-MLPA
€ 1658.00
EK20-FAM
SALSA MLPA Reagent Kit – 2000 rxn – FAM (5×6 vials)
MLPA MS-MLPA
€ 6373.00
SMR50
SALSA HhaI – 115 μl
MS-MLPA
€ 48.00

Sample DNAs (included)

A vial is included with every order of this probemix, but additional vials can also be purchased separately.

Item no.
Description
Technology
Price
SD094
SALSA Binning DNA SD094 – 6 rxn
MS-MLPA
€ 25.00

Other products

These optional accessories can be ordered separately.

Item no.
Description
Technology
Price
SMR05
SALSA FFPE Solution (SFS) – 50 rxn (10 ml)
MLPA MS-MLPA
€ 47.00

Price details & ordering

The prices above are list prices for direct orders from MRC Holland. Contact us for a quote that takes discounts and additional costs (such as shipping costs) into account. Different prices apply for orders through one of our sales partners; contact your local supplier for a quote.

More information about ordering & prices

Positive samples

Inclusion of a positive sample is usually not required, but can be useful for the analysis of your experiments. MRC Holland has very limited access to positive samples and cannot supply such samples. We recommend using positive samples from your own collection. Alternatively, you can use positive samples from an online biorepository, such as the Coriell Institute.

See this support article for commercially available positive samples that can be used with this product.

Resources

General MLPA resources

MLPA education

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