ArticleNext generation, sequencing & applications2016
Targeted Exome Sequencing Outcome Variations of Colorectal Tumors within and across Two Sequencing Platforms.
Article in Next generation, sequencing & applications, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Driver genes exome sequencing reveals distinct variants in African Americans with colorectal neoplasia.Oncotarget · 2019Article
- Species classifier choice is a key consideration when analysing low-complexity food microbiome data.Microbiome · 2018Article
- Next-generation sequencing: recent applications to the analysis of colorectal cancer.Journal of translational medicine · 2017Review
- Distinctive DNA mismatch repair and APC rare variants in African Americans with colorectal neoplasia.Oncotarget · 2017Article
- Targeted exome sequencing reveals distinct pathogenic variants in Iranians with colorectal cancer.Oncotarget · 2017Article
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Authors and funding
6 authors.
Funding
Abstract
BACKGROUND AND
aimNext generation sequencing (NGS) has quickly the tool of choice for genome and exome data generation. The multitude of sequencing platforms as well as the variabilities within each platform need to be assessed. In this paper we used two platforms (ION TORRENT AND ILLUMINA) to assess single nucleotides variants in colorectal cancer (CRC) specimens.
methodsCRC specimens (n = 13) collected from 6 CRC (cancer and matched normal) patients were used to establish the mutational profile using ION TORRENT AND ILLUMINA sequencing platforms. We analyzed a set of samples from Formalin Fixed Paraffin Embedded and FF (FF) samples on both platforms to assess the effect of sample nature (FFPE vs. FF) on sequencing outcome and to evaluate the similarity/differences of SNVs across the two platforms. In addition, duplicates of FF samples were sequenced on each platform to assess variability within platform.
resultsThe comparison of FF replicates to each other gave a concordance of 77% (± 15.3%) in Ion Torrent and 70% (± 3.7%) in Illumina. FFPE vs. FF replicates gave a concordance of 40% (± 32%) in Ion Torrent and 49% (± 19%) in Illumina. For the cross platform concordance were FFPE compared to FF (Average of 75% (± 9.8%) for FFPE samples and 67% (± 32%) for FF and 70% (± 26.8%) overall average).
conclusionOur data show a significant variability within and across platforms. Also the number of detected variants depend on the nature of the specimen; FF vs. FFPE. Validation of NGS discovered mutations is a must to rule-out false positive mutants. This validation might either be performed through a second NGS platform or through Sanger sequencing.
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