期刊
GENETIC TESTING AND MOLECULAR BIOMARKERS
卷 20, 期 2, 页码 90-93出版社
MARY ANN LIEBERT, INC
DOI: 10.1089/gtmb.2015.0229
关键词
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资金
- National Natural Science Foundation of China [81272371]
Aim: In gliomas, mutations in the core promoter region of the telomerase reverse transcriptase (TERT) gene have been associated with specific subtypes and are inversely correlated with IDH1 mutation status, predicting poor prognosis. Thus, TERT promoter mutation status might be a candidate for development as a prognostic biomarker. However, current IDH1 mutation detection methods using conventional polymerase chain reaction (PCR), followed by Sanger sequencing, have low sensitivity and are time-consuming. To improve test efficacy, we developed a more efficient detection protocol based on an amplification refractory mutation system-PCR (ARMS-PCR), which is based on the principle that DNA extension only happens when the 3'-terminal nucleotide of a primer matches its target sequence. Materials and Methods: We generated plasmids containing TERT promoter sequences and optimized this new protocol for the identification of the two most common TERT promoter mutations, C250T and C228T. Results: The enhanced sensitivity and efficiency of this protocol were validated using 124 human glioma samples. Conclusion: We have described an ARMS-PCR methodology with improved sensitivities that could replace current commonly used methods for the detection of TERT promoter mutations in gliomas.
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