| Autor: |
Corless, Broderick C., Chang, Gregory A., Cooper, Samantha, Syeda, Mahrukh M., Shao, Yongzhao, Osman, Iman, Karlin-Neumann, George, Polsky, David |
| Zdroj: |
The Journal of Molecular Diagnostics; March 2019, Vol. 21 Issue: 2 p274-285, 12p |
| Abstrakt: |
Detecting mutations in the plasma of patients with solid tumors is becoming a valuable method of diagnosing and monitoring cancer. The TERTpromoter is mutated at high frequencies in multiple cancer types, most commonly at positions -124 and -146 (designated C228Tand C250T, respectively). Detection of these mutations has been challenging because of the high GC content of this region (approximately 80%). We describe development of novel probe-based droplet digital PCR assays that specifically detect and quantify these two mutations, along with the less common 242-243 CC>TTmutation, and demonstrate their application using human tumor and plasma samples from melanoma patients. Assay designs and running conditions were optimized using cancer cell line genomic DNAs with the C228Tor C250Tmutations. The limits of detection were 0.062% and 0.051% mutant allele fraction for the C228Tand C250Tassays, respectively. Concordance of 100% was observed between droplet digital PCR and sequencing-based orthogonal methods in the detection of TERTmutant DNA in 32 formalin-fixed, paraffin-embedded melanoma tumors. TERTmutantDNA was also identified in 21 of 27 plasma samples (78%) from patients with TERTmutanttumors, with plasma mutant allele fractions ranging from 0.06% to 15.3%. There were no false positives in plasma. These data demonstrate the potential of these assays to specifically detect and quantify TERTmutantDNA in tumors and plasma of cancer patients. |
| Databáze: |
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