Jinbo Liu1,2, Zhihua Zhao2, Miaomiao Sun3, Kuisheng Chen2, Weitang Yuan1, Guozhong Jiang2. 1. 1 Department of Colorectal and Anal Surgery, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, Henan, China . 2. 2 Department of Pathology, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, China . 3. 3 Department of Pathology, Henan Tumor Hospital , Zhengzhou, China .
Abstract
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.
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 humanglioma 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.