title: The Sensitive Detection of Telomerase Reverse Transcriptase Promoter Mutation by Amplification Refractory Mutation System-PCR.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: 1 Department of Colorectal and Anal Surgery, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, Henan, China .|2 Department of Pathology, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, China .
authors.forename: Jinbo
authors.initials: J
authors.lastname: Liu
authors.affiliation: 2 Department of Pathology, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, China .
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: 3 Department of Pathology, Henan Tumor Hospital , Zhengzhou, China .
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Sun
authors.affiliation: 2 Department of Pathology, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, China .
authors.forename: Kuisheng
authors.initials: K
authors.lastname: Chen
authors.affiliation: 1 Department of Colorectal and Anal Surgery, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, Henan, China .
authors.forename: Weitang
authors.initials: W
authors.lastname: Yuan
authors.affiliation: 2 Department of Pathology, The First Affiliated Hospital of Zhengzhou University , Zhengzhou, China .
authors.forename: Guozhong
authors.initials: G
authors.lastname: Jiang
mesh_terms: D001254:Astrocytoma / Q000235:genetics*; D001932:Brain Neoplasms / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics*; D009837:Oligodendroglioma / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic; D019098:Telomerase / Q000235:genetics*
chemical_list: D009363:Neoplasm Proteins; C509186:TERT protein, human; D019098:Telomerase