https://scholars.lib.ntu.edu.tw/handle/123456789/597110
標題: | Clinical validation of KRAS, BRAF, and EGFR mutation detection using next-generation sequencing | 作者: | Lin M.-T. Mosier S.L. Thiess M. Beierl K.F. Debeljak M. LI-HUI TSENG Chen G. Yegnasubramanian S. Ho H. Cope L. Wheelan S.J. Gocke C.D. Eshleman J.R. |
關鍵字: | BRAF; Deamination; EGFR; KRAS; Next-generation sequencing; Read depth; Validation | 公開日期: | 2014 | 出版社: | American Society of Clinical Pathologists | 卷: | 141 | 期: | 6 | 起(迄)頁: | 856-866 | 來源出版物: | American Journal of Clinical Pathology | 摘要: | Objectives: To validate next-generation sequencing (NGS) technology for clinical diagnosis and to determine appropriate read depth. Methods: We validated the KRAS, BRAF, and EGFR genes within the Ion AmpliSeq Cancer Hotspot Panel using the Ion Torrent Personal Genome Machine (Life Technologies, Carlsbad, CA). Results: We developed a statistical model to determine the read depth needed for a given percent tumor cellularity and number of functional genomes. Bottlenecking can result from too few input genomes. By using 16 formalin-fixed, paraffin-embedded (FFPE) cancer-free specimens and 118 cancer specimens with known mutation status, we validated the six traditional analytic performance characteristics recommended by the Next-Generation Sequencing: Standardization of Clinical Testing Working Group. Baseline noise is consistent with spontaneous and FFPE-induced C:G→T:A deamination mutations. Conclusions: Redundant bioinformatic pipelines are essential, since a single analysis pipeline gave false-negative and false-positive results. NGS is sufficiently robust for the clinical detection of gene mutations, with attention to potential artifacts. ? American Society for Clinical Pathology. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-84904502195&doi=10.1309%2fAJCPMWGWGO34EGOD&partnerID=40&md5=30eb3683a4b8dd5a5bd4911ed44a5d94 https://scholars.lib.ntu.edu.tw/handle/123456789/597110 |
ISSN: | 0002-9173 | DOI: | 10.1309/AJCPMWGWGO34EGOD | SDG/關鍵字: | B Raf kinase; epidermal growth factor receptor; formaldehyde; K ras protein; paraffin; article; bottleneck population; gene mutation; gene sequence; genetic procedures; genome; genotype; human; human cell; human tissue; major clinical study; next generation sequencing; point mutation; priority journal; pyrosequencing; single nucleotide polymorphism; BRAF; Deamination; EGFR; KRAS; Next-generation sequencing; Read depth; Validation; Deamination; High-Throughput Nucleotide Sequencing; Limit of Detection; Molecular Diagnostic Techniques; Mutation; Neoplasms; Paraffin Embedding; Proto-Oncogene Proteins; Proto-Oncogene Proteins B-raf; ras Proteins; Receptor, Epidermal Growth Factor; Reproducibility of Results; Sensitivity and Specificity; Tumor Markers, Biological |
顯示於: | 基因體暨蛋白體醫學研究所 |
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