A Systematic Assessment of Accuracy in Detecting Somatic Mosaic Variants by Deep Amplicon Sequencing: Application to NF2 Gene

摘要: The accurate detection of low-allelic variants is still challenging, particularly for the identification somatic mosaicism, where matched control sample not available. High throughput sequencing, by simultaneous and independent analysis thousands different DNA fragments, might overcome many limits traditional methods, greatly increasing sensitivity. However, it necessary to take into account high number false positives that may arise due lack samples. Here, we applied deep amplicon sequencing samples with known genotype variant allele fraction (VAF) followed a tailored statistical analysis. This method allowed define minimum value VAF detecting mosaic accuracy. Then, exploited estimated select candidate alterations in NF2 gene 34 unknown (30 blood 4 tumor DNAs), demonstrating suitability our method. strategy propose optimizes use low abundance variants. Moreover, can be approaches estimate background noise accuracy experimental design.