Molecular diagnostic profiling of lung cancer specimens with a semiconductor-based massive parallel sequencing approach: feasibility, costs, and performance compared with conventional sequencing.

摘要: In the context of personalized oncology, screening for somatic tumor mutations is crucial prediction an individual patient's response to therapy. Massive parallel sequencing (MPS) has been suggested routine diagnostics, but this technology not sufficiently evaluated with respect feasibility, reliability, and cost effectiveness diagnostic formalin-fixed, paraffin-embedded material. We performed ultradeep targeted semiconductor-based MPS (190 amplicons covering hotspot in 46 genes) a variety samples lung adenocarcinoma tissue known EGFR (n = 28). The reflected typical spectrum material including small biopsies low tumor-cell content. Using MPS, we successfully sequenced all samples, mean read depth 2947 reads per amplicon. High-quality sequence were obtained from containing ≥10% one sample, variant calling identified same as detected by conventional Sanger sequencing. Moreover, 43 additional 17 genes amplifications ERBB2 genes. performance was reliable independent type material, well fixation extraction methods, influenced content degree DNA degradation. sample multiplexing, focused approached diagnostically acceptable rates.