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SALSA MLPA Probemix P088 Oligodendroglioma 1p-19q

Oligodendroglioma

Region: 1p, 19q, IDH1 (2q33.3), IDH2 (15q26.1) and CDKN2A/2B 9p21.3

MLPA | CE
Intended purpose
The SALSA MLPA Probemix P088 Oligodendroglioma 1p-19q is an in vitro diagnostic (IVD)1 or research use only (RUO) semi-quantitative assay2 for the detection of co-deletion of chromosome arms 1p and 19q, and deletions of the CDKN2A and CDKN2B genes, and for detection of the most common somatic point mutations in IDH1 (p.R132H and p.R132C) and IDH2 (p.R172K and p.R172M) in genomic DNA isolated from fresh-frozen or formalin-fixed paraffin-embedded (FFPE) human glioma specimens. P088 Oligodendroglioma 1p-19q is intended to aid in diagnosis of oligodendroglioma.

Deletions of CDKN2A and/or CDKN2B detected with P088 Oligodendroglioma 1p-19q should be confirmed with a different technique. In particular, deletions detected by only a single probe always require confirmation by another method. Except for the four mutations mentioned above, no other mutations in IDH1 and IDH2 can be detected with P088 Oligodendroglioma 1p-19q. For detection of rare mutations in IDH1 and IDH2, it is recommended to use this assay in combination with sequence analysis.

Assay results are intended to be used in conjunction with other clinical and diagnostic findings, consistent with professional standards of practice, including confirmation by alternative methods. The results of this test should be interpreted by a clinical molecular geneticist or equivalent.

1Please note that this probemix is for in vitro diagnostic (IVD) use in the countries specified at the end of this product description. In all other countries, the product is for research use only (RUO).
2To be used in combination with a SALSA MLPA Reagent Kit, Coffalyser.Net analysis software, and SALSA Binning DNA SD079.

Clinical background
Oligodendrogliomas are a type of glioma (central nervous system neoplasm) that originate from the oligodendrocytes of the brain or from a glial precursor cell. According to the WHO Classification of Tumours of the Central Nervous System, oligodendroglioma is molecularly defined by co-deletion of chromosome arms 1p and 19q in combination with a mutation in IDH1 or IDH2 (Louis et al. 2016; Louis et al. 2021). The 1p/19q co-deletion distinguishes oligodendrogliomas from other gliomas, and is considered an early event in oligodendroglioma tumorigenesis (Pinkham et al. 2015). The co-deletion is thought to be the result of an unbalanced whole-arm translocation between chromosomes 1 and 19 with the loss of the resulting hybrid chromosome (Griffin et al. 2006; Jenkins et al. 2006). The IDH1 p.R132H mutation accounts for ~90% of all IDH1 and IDH2 mutations. The remaining ~10% are other substitutions in codon 132 of IDH1 (e.g. p.R132C, p.R132S, p.R132G) or substitutions in codon 172 of IDH2 (p.R172K, p.R172M or p.R172W) (Cahill et al. 2015; Hartmann et al. 2009). p.R132H accounts for 92.7% and p.R132C for 4% of all IDH1 mutations, and p.R172K accounts for 64.5% and p.R172M for 19% of all IDH2 mutations (Hartmann et al. 2009).

Probemix content
The SALSA MLPA Probemix P088-D1 Oligodendroglioma 1p-19q contains 59 MLPA probes with amplification products between 126 and 509 nucleotides (nt). This includes 19 probes for the 1p arm and 12 probes for the 19q arm. In addition, three flanking probes for the 1q arm and two flanking probes for the 19p arm have been included to discriminate between chromosome arm losses and whole chromosome losses. The probemix also contains four probes specific for the p.R132H and p.R132C mutations in IDH1 and the p.R172K and p.R172M mutations in IDH2 which will only generate a signal when the mutation is present. Furthermore, the probemix contains three probes for CDKN2A and two probes for CDKN2B. Finally, 14 reference probes are included that target relatively copy number stable regions in central nervous system tumours, especially in oligodendrogliomas. Complete probe sequences and the identity of the genes detected by the reference probes are available online (www.mrcholland.com) and in Table 3, respectively.

This probemix contains nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at www.mrcholland.com.

SALSA Binning DNA SD079
The Binning DNA SD079 provided with this probemix can be used for binning of all probes including the four mutation-specific probes (203 nt probe 19529-L16492 for the IDH1 p.R132H (c.395G>A) mutation, 227 nt probe 14787-L23353 for the IDH1 p.R132C (c.394C>T) mutation, 238 nt probe 20963-L29002 for the IDH2 p.R172K (c.515G>A) mutation, and 244 nt probe 20963-L29001 for the IDH2 p.R172M (c.515G>T) mutation). Binning DNA SD079 is a mixture of genomic DNA from healthy individuals and synthetic DNA that contains the target sequences detected by the above mentioned probes. Inclusion of one reaction with 5 μl Binning DNA SD079 in initial MLPA experiments is essential as it can be used to aid in data binning of the peak pattern using Coffalyser.Net software. Furthermore, Binning DNA should be included in the experiment whenever changes have been applied to the set-up of the capillary electrophoresis device (e.g. when capillaries have been renewed). Binning DNA should never be used as a reference sample in the MLPA data analysis, neither should it be used in quantification of mutation signal(s). It is strongly advised that all samples tested are extracted with the same method and derived from the same source of tissue. For further details, please consult the Binning DNA SD079 product description, available online: www.mrcholland.com.

Sample DNA
Sample DNA developed for this product:

Order Items

Probemix

Item no.
Description
Technology
Price
P088-025R
SALSA MLPA Probemix P088 Oligodendroglioma 1p-19q – 25 rxn
€ 281.00
P088-050R
SALSA MLPA Probemix P088 Oligodendroglioma 1p-19q – 50 rxn
€ 550.00
P088-100R
SALSA MLPA Probemix P088 Oligodendroglioma 1p-19q – 100 rxn
€ 1075.00

Required Reagents (Sold Separately)

Item no.
Description
Technology
Price
EK1-FAM
SALSA MLPA Reagent Kit – 100 rxn – FAM
€ 341.00
EK1-Cy5
SALSA MLPA Reagent Kit – 100 rxn – Cy5
€ 341.00
EK5-FAM
SALSA MLPA Reagent Kit – 500 rxn – FAM
€ 1571.00
EK5-Cy5
SALSA MLPA Reagent Kit – 500 rxn – Cy5
€ 1571.00
EK20-FAM
SALSA MLPA Reagent Kit – 2000 rxn – FAM
€ 6037.00

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CE

CE-marked products are for In Vitro Diagnostic (IVD) use only in EU (candidate) member states and members of the European Free Trade Association (EFTA), and the UK.