ERASE-Seq variant caller- superior performance to 0.1% allele frequency and below, without barcodes!

Finally, the promise of sensitive, accurate liquid biopsy analysis by NGS has been realized! ERASE-Seq uses inter-sample signal processing to eliminate background artifacts and random sequencing errors, all without the need for molecular barcodes. ERASE-Seq has been proven in sequencing of ctDNA and CTC samples, and excels in the allele frequency range of 0.05% to 0.5%, where barcode approaches lose both sensitivity and specificity.  

How Does ERASE-Seq Work?

ERASE-Seq (Elimination of Recurrent Artifacts and Stochastic Errors) functions by quantitatively modeling the background error profile of normal controls that have gone through the library prep and sequencing process in order to establish the inherent noise distribution for each possible variant. The background modeling is done once and then embedded in the ERASE-seq algorithm. Variant calls are then made by comparing sample sequencing data to the background error distribution, providing large increases in both sensitivity and specificity. To further improve detection sensitivity, samples can be split into two or more reactions to allow statistical analysis between the replicates as well as to the background model.

The use of this inter-sample statistical analysis, as opposed to the more typical intra-sample QC metrics, provides substantial performance gains. This approach eliminates both random errors and systematic biases inherent in all sequencing workflows. ERASE-Seq delivers superior detection sensitivity to 0.05% allele frequency (AF), with a false positive rate 10-100X lower than leading molecular barcode approaches1. ERASE-Seq analysis is accessed by uploading raw fastq files via secure upload to a HIPAA-compliant cloud-based server. VCF and Excel variant reports are available for download within 24 hours. Local software installations are also available.

ERASE-Seq is modular and flexible, and can be tailored based on desired performance, sequencing depth, and available DNA:

Standard Background Correction Only Ultra-sensitive
90% Sensitivity LOD (AF) 0.1-0.2% 0.25-0.5% 0.05-0.1%
False Positives at LOD 0.1 0.1 0.1
DNA Input Requirement 20ng 10ng 30ng
Overall Sequencing Depth 15,000X 7,500X 30,000X

ERASE-Seq is the proven solution for NGS of liquid biopsy samples. Key benefits include:

  • Superior sensitivity and 10-100X fewer false positives than molecular barcodes (UMIs)

  • Excels in the 0.05%-0.5% AF range where molecular barcodes struggle

  • Any targeted panel can be adapted easily to ERASE-Seq with software validation- and no barcodes needed

  • Provides similar detection performance to digital PCR, but ERASE-Seq can test for thousands of variants per test

  • Customized variant target lists

  • Validated in numerous sequencing labs globally

ERASE-Seq Purchase Options

  1. Purchase a software license to run ERASE-Seq on a validated panel that you source elsewhere (e.g., Illumina Trusight Tumor 15 or Swift 56G).

  2. ERASE-Seq is included in the cost of any panels you purchase from Fluxion, including our standard Spotlight 59 pan-cancer panel and custom panels we supply.

ERASE-Seq can be applied to virtually any targeted sequencing panel. Currently-validated panels include Fluxion's own Spotlight 59, Illumina Trusight Tumor 15, and Swift 56G. Additional ERASE-Seq technical information and IFUs are available below and in our Library.

Custom Panel Development

Fluxion offers customized Spotlight Panels optimized to enable fast, comprehensive targeted sequencing. Leveraging a combination of unique design and chemistry, custom Spotlight Panels produce overlapping (tiled) amplicons in a single-tube using a rapid 2-hour workflow. Combined with ERASE-Seq bioinformatics, this unique approach provides a powerful solution for detecting ultra-low frequency somatic mutations using low input DNA from challenging samples types such ctDNA. A fast and easy single-tube workflow produces the best-in-class performance for on-target percentage and coverage uniformity, enabling low frequency variant discovery and confirmation. To get started with a custom Spotlight panel, fill out the custom panel design template and submit to info@fluxionbio.com.