Research Papers:

Validation and optimization of the Ion Torrent S5 XL sequencer and Oncomine workflow for BRCA1 and BRCA2 genetic testing

Saeam Shin, Yoonjung Kim, Seoung Chul Oh, Nae Yu, Seung-Tae Lee, Jong Rak Choi and Kyung-A Lee _

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Oncotarget. 2017; 8:34858-34866. https://doi.org/10.18632/oncotarget.16799

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Saeam Shin1,2,*, Yoonjung Kim1,*, Seoung Chul Oh1, Nae Yu1, Seung-Tae Lee1, Jong Rak Choi1, Kyung-A Lee1

1Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Republic of Korea

2Department of Laboratory Medicine, Hallym University College of Medicine, Kangnam Sacred Heart Hospital, Seoul, Republic of Korea

*These authors contributed equally to this work

Correspondence to:

Kyung-A Lee, email: [email protected]

Keywords: BRCA, Ion Torrent, next-generation sequencing, S5 XL, Oncomine

Received: December 22, 2016     Accepted: March 24, 2017     Published: April 03, 2017


In this study, we validated the analytical performance of BRCA1/2 sequencing using Ion Torrent’s new bench-top sequencer with amplicon panel with optimized bioinformatics pipelines. Using 43 samples that were previously validated by Illumina’s MiSeq platform and/or by Sanger sequencing/multiplex ligation-dependent probe amplification, we amplified the target with the Oncomine BRCA Research Assay and sequenced on Ion Torrent S5 XL (Thermo Fisher Scientific, Waltham, MA, USA). We compared two bioinformatics pipelines for optimal processing of S5 XL sequence data: the Torrent Suite with a plug-in Torrent Variant Caller (Thermo Fisher Scientific), and commercial NextGENe software (Softgenetics, State College, PA, USA). All expected 681 single nucleotide variants, 15 small indels, and three copy number variants were correctly called, except one common variant adjacent to a rare variant on the primer-binding site. The sensitivity, specificity, false positive rate, and accuracy for detection of single nucleotide variant and small indels of S5 XL sequencing were 99.85%, 100%, 0%, and 99.99% for the Torrent Variant Caller and 99.85%, 99.99%, 0.14%, and 99.99% for NextGENe, respectively. The reproducibility of variant calling was 100%, and the precision of variant frequency also showed good performance with coefficients of variation between 0.32 and 5.29%. We obtained highly accurate data through uniform and sufficient coverage depth over all target regions and through optimization of the bioinformatics pipeline. We confirmed that our platform is accurate and practical for diagnostic BRCA1/2 testing in a clinical laboratory.

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