Identification of single nucleotide variants using position-specific error estimation in deep sequencing data | BMC Medical Genomics | Full Text

Identification of single nucleotide variants using position-specific error estimation in deep sequencing data | BMC Medical Genomics | Full Text

BMC Medical Genomics

Identification of single nucleotide variants using position-specific error estimation in deep sequencing data

• Dimitrios Kleftogiannis, 
• Marco Punta, 
• Anuradha Jayaram, 
• Shahneen Sandhu, 
• Stephen Q. Wong, 
• Delila Gasi Tandefelt, 
• Vincenza Conteduca, 
• Daniel Wetterskog, 
• Gerhardt Attard & 
• Stefano Lise 

BMC Medical Genomicsvolume 12, Article number: 115 (2019) | Download Citation

Abstract

Background

Targeted deep sequencing is a highly effective technology to identify known and novel single nucleotide variants (SNVs) with many applications in translational medicine, disease monitoring and cancer profiling. However, identification of SNVs using deep sequencing data is a challenging computational problem as different sequencing artifacts limit the analytical sensitivity of SNV detection, especially at low variant allele frequencies (VAFs).

Methods

To address the problem of relatively high noise levels in amplicon-based deep sequencing data (e.g. with the Ion AmpliSeq technology) in the context of SNV calling, we have developed a new bioinformatics tool called AmpliSolve. AmpliSolve uses a set of normal samples to model position-specific, strand-specific and nucleotide-specific background artifacts (noise), and deploys a Poisson model-based statistical framework for SNV detection.

Results

Our tests on both synthetic and real data indicate that AmpliSolve achieves a good trade-off between precision and sensitivity, even at VAF below 5% and as low as 1%. We further validate AmpliSolve by applying it to the detection of SNVs in 96 circulating tumor DNA samples at three clinically relevant genomic positions and compare the results to digital droplet PCR experiments.

Conclusions

AmpliSolve is a new tool for in-silico estimation of background noise and for detection of low frequency SNVs in targeted deep sequencing data. Although AmpliSolve has been specifically designed for and tested on amplicon-based libraries sequenced with the Ion Torrent platform it can, in principle, be applied to other sequencing platforms as well. AmpliSolve is freely available at https://github.com/dkleftogi/AmpliSolve.