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Article

Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing

  • Authors:
    • Juan Li
    • Haiyan Zhang
    • Li Wang
    • Chuanhong Yang
    • Huangwen Lai
    • Wei Zhang
    • Xiaodong Chen
    • Jie Wang
  • View Affiliations / Copyright

    Affiliations: Department of Biochemistry and Molecular Biology, School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, Guangdong 510006, P.R. China, Department of Medical Research, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou, Guangdong 510010, P.R. China, Department of Pathology, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou, Guangdong 510010, P.R. China
  • Pages: 4376-4381
    |
    Published online on: June 23, 2015
       https://doi.org/10.3892/mmr.2015.3987
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Abstract

Patients with glioblastomas with a specific mutation in the isocitrate dehydrogenase 1 (IDH1) gene have a better prognosis than those with gliomas with wild‑type IDH1. IDH1 analysis has become part of the standard diagnostic procedure and a promising tool used for stratification in clinical trials. The present study aimed to compare high resolution melting (HRM) analysis, immunohistochemistry (IHC) and direct DNA sequencing for the detection of IDH mutations in gliomas. Fifty‑one formalin‑fixed paraffin‑embedded tumor samples were selected. For the HRM analysis and direct DNA sequencing, DNA was extracted from the tissues. For IHC, sections were stained with an anti‑IDH1‑R132H specific antibody. The HRM analysis method identified 33 cases of IDH1 gene mutations, and all mutations occurred at the R132H site. There were 33 cases of IDH1 gene mutations found by IHC, which was consistent with that identified using the HRM analysis method. However, only 30 IDH1 samples were confirmed by sequencing, in which mutations occurred at the IDH1 exon 4 R132H site. No mutation was detected in the other three of these 33 cases (two grade II oligodendroglioma and one grade II diffuse astrocytoma) by sequencing, while IHC was positive for IDH1‑R132H. The results showed that the mutation detection rate was not identified to be significantly different (P=0.250) when determined by the HRM analysis method or by direct DNA sequencing, as the concordant rate between the two methods was high (κ=0.866). The HRM analysis method in glioma IDH1 gene mutation detection has advantages of high sensitivity, good repeatability, simple operation and accurate results. It provides a novel method for detecting mutations of the IDH1 gene in paraffin embedded tissue samples of clinical glioma. Related to a small amount of sample, there was no evidence showing that HRM analysis method is superior to IHC. Direct DNA sequencing, HRM analysis and IHC results were consistent; however, HRM and IHC are more sensitive than direct DNA sequencing in identifying the IDH1‑R132H mutation.
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Copy and paste a formatted citation
Spandidos Publications style
Li J, Zhang H, Wang L, Yang C, Lai H, Zhang W, Chen X and Wang J: Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing. Mol Med Rep 12: 4376-4381, 2015.
APA
Li, J., Zhang, H., Wang, L., Yang, C., Lai, H., Zhang, W. ... Wang, J. (2015). Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing. Molecular Medicine Reports, 12, 4376-4381. https://doi.org/10.3892/mmr.2015.3987
MLA
Li, J., Zhang, H., Wang, L., Yang, C., Lai, H., Zhang, W., Chen, X., Wang, J."Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing". Molecular Medicine Reports 12.3 (2015): 4376-4381.
Chicago
Li, J., Zhang, H., Wang, L., Yang, C., Lai, H., Zhang, W., Chen, X., Wang, J."Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing". Molecular Medicine Reports 12, no. 3 (2015): 4376-4381. https://doi.org/10.3892/mmr.2015.3987
Copy and paste a formatted citation
x
Spandidos Publications style
Li J, Zhang H, Wang L, Yang C, Lai H, Zhang W, Chen X and Wang J: Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing. Mol Med Rep 12: 4376-4381, 2015.
APA
Li, J., Zhang, H., Wang, L., Yang, C., Lai, H., Zhang, W. ... Wang, J. (2015). Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing. Molecular Medicine Reports, 12, 4376-4381. https://doi.org/10.3892/mmr.2015.3987
MLA
Li, J., Zhang, H., Wang, L., Yang, C., Lai, H., Zhang, W., Chen, X., Wang, J."Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing". Molecular Medicine Reports 12.3 (2015): 4376-4381.
Chicago
Li, J., Zhang, H., Wang, L., Yang, C., Lai, H., Zhang, W., Chen, X., Wang, J."Comparative study of IDH1 mutations in gliomas by high resolution melting analysis, immunohistochemistry and direct DNA sequencing". Molecular Medicine Reports 12, no. 3 (2015): 4376-4381. https://doi.org/10.3892/mmr.2015.3987
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