Improved Efficiency and Reliability of NGS Amplicon Sequencing Data Analysis for Genetic Diagnostic Procedures Using AGSA Software

Figure 3

AGSA detected efficiently all the variants reported in Sanger analysis. Efficiencies of Sanger sequencing versus Roche pyrosequencing analysed with AGSA. The blue bars represent confirmed variants. The green bars represent false positive variants (technical artefacts). For pyrosequencing, false positives are defined as variants not confirmed by Sanger sequencing. For Sanger sequencing, false positives were not found by pyrosequencing and were not confirmed by a second Sanger run of the same sample. The red bars represent false negative variants. No false negative was found by pyrosequencing. False negatives for Sanger analysis were detected by pyrosequencing and they were actually found on a second Sanger run of the same sample. The yellow bar represents variants that were not called by AGSA because of poor coverage (inducing a number of variant reads < 4).