HaloPlex Targeted Resequencing for Mutation Detection in Clinical Formalin-Fixed, Paraffin-Embedded Tumor Samples.

摘要: In recent years, the advent of massively parallel next-generation sequencing technologies has enabled substantial advances in study human diseases. Combined with targeted DNA enrichment methods, high sequence coverage can be obtained for different genes simultaneously at a reduced cost per sample, creating unique opportunities clinical cancer diagnostics. However, formalin-fixed, paraffin-embedded (FFPE) process tissue samples, routinely used pathology departments, results fragmentation and nucleotide modifications that introduce number technical challenges downstream biomolecular analyses. We evaluated HaloPlex target system somatic mutation detection 80 fractions derived from 20 cases paired tumor normal available both FFPE fresh-frozen format. Several to standard method were introduced, including fragment length two strand capturing. found material HaloPlex-based sequencing, even when starting small amounts DNA. By specifically capturing strands each fragment, we able reduce false-positive errors caused by FFPE-induced artifacts lower limit mutations. believe presented here will broadly applicable as tool settings.