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| Preliminary Establishment of High Resolution Melting Analysis for Detecting IDH2 Gene Mutations in Gliomas |
| WANG Yin-hui1, ZHANG Hai-yan1, YANG Chuan-hong1, ZHANG Wei2, LAI Huang-wen1, CHEN Xiao-dong2, WANG Jie2 |
1. Department of Medical Research, General Hospital of Guangzhou Military Command, Guangzhou 510010, China;
2. Department of Pathology, General Hospital of Guangzhou Military Command, Guangzhou 510010, China |
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Abstract Isocitrate dehydrogenase2(IDH2) mutations have recently been identified as early and frequent genetic alterations in astrocytomas, oligodendrogliomas, and oligoastrocytomas, as well as secondary glioblastomas. Since the mutations have been associated with better clinical outcome, they have shown diagnostic and prognostic value. It was performed with optimizing conditions to preliminary establish a high resolution melting for the detecting of IDH2 mutations in routine formalin-fixed tissues of brain biopsies. IDH2 mutations in paraffin-embedded tissues of gliomas were detected by HRM and direct sequencing, estimating the stability and sensitivity of reaction system and reaction conditions. The optimal annealing temperature was 52.5 ℃. The optimization of 20 μl-HRM reaction system including 0.6 μmol/L primer, 45 ng DNA template,2.75 mmol/L Mg2+, was established by response surface Design. The results were consistent with the direct sequencing but have a higher sensitivity. It showed that HRM was simple, specific, accurate and reliable method. It would provide the reference for the clinical application of detection IDH2 mutations.
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Received: 24 September 2013
Published: 25 December 2013
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