Prognostic Value of IDH1 Mutations Identified with PCR-RFLP Assay in Glioblastoma Patients

Bujko, Mateusz; Kober, Paulina; Matyja, Ewa; Nauman, Paweł; Dyttus-Cebulok, Katarzyna; Czeremszynska, B.; Bonicki, Wiesław; Siedlecki, Janusz A.

doi:10.1007/bf03256369

Cited by 25 publications

(24 citation statements)

References 26 publications

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“…Bujko et al 60 and Meyer et al 61 have adapted PCR-based restriction fragment length polymorphism assays to detect mutations in codon 132 of the IDH1 gene. These assays, also known as derived cleaved amplified polymorphic sequence analysis, are based on the presence or absence of a restriction enzyme binding site at codon R132 in IDH1 , depending on the presence or absence of mutation.…”

Section: Detection Of Idh Mutationsmentioning

confidence: 99%

“…Identifying the specific IDH1 mutation present requires additional assays using multiple PCR primer pairs and restriction enzymes and is therefore quite labour intensive. However, the tests are relatively rapid and have the advantage of not requiring expensive sequencing equipment 60 61. We have found this technique useful for screening purposes (mutant vs wild type in antibody-negative cases; figure 2B,C), and for the confirmation of equivocal sequencing results.…”

Section: Detection Of Idh Mutationsmentioning

confidence: 99%

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Isocitrate dehydrogenase mutations in diffuse gliomas: clinical and aetiological implications

et al. 2011

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The discovery of isocitrate dehydrogenase (IDH) mutations in gliomas is one example of the large impact that next-generation sequencing is having on the understanding of tumour biology and human disease in general. IDH mutations are early and common events in the development of astrocytomas, oligodendrogliomas and oligoastrocytomas. IDH mutations are also found in some myeloid malignancies and soft tissue tumours, but are rare in other malignancies. IDH mutation detection can be incorporated into routine pathology practice via immunohistochemistry and/or standard sequencing techniques and has great diagnostic value. An emerging theme is that IDH mutation status in gliomas is of great prognostic relevance, and there are proposals to include IDH mutation status in the next iteration of the WHO classification of gliomas. The mechanisms of action(s) of mutant IDH are not fully understood, but the understanding is progressing rapidly, and may provide a mechanism to link diverse proneoplastic processes such as oxidative damage and epigenetic dysregulation. There are exciting prospects of novel therapies for glioma patients emerging from the elucidation of these mechanisms. Given the diagnostic and prognostic implications of IDH mutation, and the potential for new therapies, all gliomas should be assessed for IDH mutation status in the future.

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Section: Detection Of Idh Mutationsmentioning

confidence: 99%

Section: Detection Of Idh Mutationsmentioning

confidence: 99%

Isocitrate dehydrogenase mutations in diffuse gliomas: clinical and aetiological implications

et al. 2011

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“…A number of methods have been developed to detect IDH1 mutations including DNA sequencing (4,17), PCR-restriction fragment length polymorphism (RFLP) (18,19), PCR-single strand conformation polymorphism (SSCP) (20), pyrosequencing (21,22), and high-resolution melting curve analysis (HRM) (23,24). All of these methods have limitations in terms of sensitivity, ease of use and throughput.…”

Section: Comparison Of Immunohistochemistry Dna Sequencing and Allelmentioning

confidence: 99%

Comparison of immunohistochemistry, DNA sequencing and allele-specific PCR for the detection of IDH1 mutations in gliomas

et al. 2012

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Abstract. Previous studies have identified mutations of the isocitrate dehydrogenase 1 (IDH1) gene in more than 70% of World Health Organization (WHO) grade II and III gliomas. The most frequent mutation leads to a specific amino acid change from arginine to histidine at codon 132 (c.395G>A, p.R132H). IDH1 mutated tumors have a better prognosis than IDH1 non-mutated tumors. The aim of our study was to evaluate and compare the methods of mIDH1 R132H immunohistochemistry, allele-specific PCR and DNA sequencing for determination of IDH1 status. We performed a retrospective study of 91 patients with WHO grade II (n=43) and III (n=48) oligodendrogliomas. A fragment of exon 4 spanning the sequence encoding the catalytic domain of IDH1, including codon 132, was amplified and sequenced using standard conditions. Allele-specific amplification was performed using two forward primers with variations in their 3' nucleotides such that each was specific for the wild-type or the mutated variant, and one reverse primer. Immunohistochemistry was performed with mouse monoclonal mIDH1 R132H. DNA was extracted from FFPE sections following macrodissection. IDH1 mutations were found in 55/90 patients (61.1%) by direct sequencing. R132H mutations were found in 47/55 patients (85.4%). The results of the allele-specific PCR positively correlated with those from DNA sequencing. Other mutations (p.R132C, p.R132S and pR132G) were found by DNA sequencing in 3, 3 and 2 tumors, respectively (8/55 patients, 14.6%). mIDH1 R132H immunostaining was found in the 47 patients presenting the R132H mutation (sensitivity 47/47, 100% for this mutation). None of the tumors presenting a wild-type IDH1 gene were stained (specificity 35/35, 100%). Our results demonstrate that immunohistochemistry using the mIDH1 R132H antibody and allele-specific amplification are highly sensitive techniques to detect the most frequent mutation of the IDH1 gene.

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“…Various studies have identified candidate prognosis factors for GBM (Bujko et al, 2010; Burger and Green, 1987; Davis et al, 1949; Ohgaki et al, 2004; Stupp et al, 2005), yet it has remained unclear how best to account for the joint effects of multiple factors. In an attempt to address this problem, researchers have proposed multivariate prognosis models (Colman et al, 2010; Lamborn et al, 2004; Liang et al, 2005; Nutt et al, 2003), which attempt to account for multiple effects simultaneously.…”

Section: Introductionmentioning

confidence: 99%

Clinical and molecular models of Glioblastoma multiforme survival

Piccolo¹,

Frey²

2013

IJDMB

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Glioblastoma multiforme (GBM), a highly aggressive form of brain cancer, results in a median survival of 12–15 months. For decades, researchers have explored the effects of clinical and molecular factors on this disease and have identified several candidate prognostic markers. In this study, we evaluated the use of multivariate classification models for differentiating between subsets of patients who survive a relatively long or short time. Data for this study came from The Cancer Genome Atlas (TCGA), a public repository containing clinical, treatment, histological and biomolecular variables for hundreds of patients. We applied variable-selection and classification algorithms in a cross-validated design and observed that predictive performance of the resulting models varied substantially across the algorithms and categories of data. The best-performing models were based on age, treatments and global DNA methylation. In this paper, we summarise our findings, discuss lessons learned in analysing TCGA data and offer recommendations for performing such analyses.

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Prognostic Value of IDH1 Mutations Identified with PCR-RFLP Assay in Glioblastoma Patients

Abstract: Our results indicate that the IDH1 R132H mutation is a powerful prognostic marker in GBM treated with chemoradiation. The PCR-RFLP method allows for a fast, inexpensive, and sensitive mutation screening.

Cited by 25 publications

References 26 publications

Isocitrate dehydrogenase mutations in diffuse gliomas: clinical and aetiological implications

Isocitrate dehydrogenase mutations in diffuse gliomas: clinical and aetiological implications

Comparison of immunohistochemistry, DNA sequencing and allele-specific PCR for the detection of IDH1 mutations in gliomas

Clinical and molecular models of Glioblastoma multiforme survival

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