AI Article Synopsis
- Next-generation sequencing (NGS) methods can have issues with throughput, cost, or accuracy for targeted DNA sequencing, which is critical for mutation detection.
- A study assessed the performance of two NGS platforms (SOLiD 4 and Ion Torrent PGM) against Sanger sequencing for the BRCA1 and BRCA2 genes, reporting high sensitivity (97.8% for SOLiD and 98.9% for PGM) and specificity (100% for SOLiD).
- The workflow was found to be cost-effective (potentially under $200), requires minimal DNA, and could efficiently use DNA from both blood and buccal wash samples for clinical mutation screening.
Article Abstract
In a clinical setting, next-generation sequencing (NGS) approaches for the enrichment and resequencing of DNA targets may have limitations in throughput, cost, or accuracy. We evaluated an NGS workflow for targeted DNA sequencing for mutation detection. Targeted sequence data of the BRCA1 and BRCA2 genes, generated using a PCR-based, multiplexed NGS approach using the SOLiD 4 (n = 24) and Ion Torrent PGM (n = 20) next-generation sequencers, were evaluated against sequence data obtained by Sanger sequencing. The overall sensitivity for SOLiD and PGM were 97.8% (95% CI = 94.7 to 100.0) and 98.9% (95% CI = 96.8 to 100.0) respectively. The specificity for the SOLiD platform was high, at 100.0% (95% CI = 99.3 to 100.0). PGM correctly identified all 3 indels, but 68 false-positive indels were also called. Equimolar normalization of amplicons was not necessary for successful NGS. Both platforms are highly amenable to scale-up, potentially reducing the reagent cost for BRCA testing to
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http://dx.doi.org/10.1016/j.jmoldx.2012.06.003 DOI Listing Publication Analysis
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