This is a platform agnostic hybrid in silico proficiency testing and validated materials program for laboratories performing targeted NGS of cancer genes or mutational hotspots in hematologic malignancies.

For panel-specific hematologic malignancies bioinformatic proficiency testing challenges, refer to the NGSB3 program.

Minimum Requirements:

• Laboratories must provide a gene panel sequencing data file (FASTQ or unaligned BAM) that has been generated using their current clinical sequencing protocols from one of the following sources: a specimen from the NGS - Germline program or from one of the following NIST Reference Material cell lines: RM 8398 (NA12878), RM 8391, RM 8392, or RM 8393. FASTQs or unaligned BAMs must be submitted along with a BED file describing the regions targeted and interrogated by your laboratory. Specimens from the NGSST and NGSHM programs or additional Coriell/NIST Reference Material cell lines cannot be used for this program.
• Laboratories can transfer and download files from most modern browsers/operating systems. Due to the extremely large file sizes, 40 Mbps transfer speed or higher is needed to ensure successful transfer of your laboratory’s sequencing files to the CAP. For the most up-to-date information on system requirements, click Browser and Operating System Requirements located at the bottom of the cap.org homepage.

Additional Information, Proficiency Testing Program

• Laboratories will be asked to identify somatic single nucleotide variants and small (1-15bp) insertions, deletions, duplications, and deletions-insertions (delins) in a subset of hematologic malignancies mutational hotspots/genes with VAF potentially as low as 5%. Laboratories will be required to submit results of the variants identified.

Additional Information, Validated Materials

• The sequencing file will contain up to 75 custom somatic variants that are tailored to the specific assay submitted (depending on the size of the panel provided) at VAF from 3% to 99% (higher allele fractions to mimic loss of heterozygosity or homozygosity) and will include:


• Single nucleotide variants
• Insertions, deletions, delins, and/or duplications ranging from 1-100bp (1-15bp, 16-50bp, 51-100bp)


• All variants will be modeled based on actual somatic mutations from the COSMIC database. This portion of the program is not traditional proficiency testing and no results will be returned to the CAP; information regarding the variants introduced will be sent along with the mutagenized file.

Program Information

• The proficiency testing portion of this program is designed to complement and augment NGS somatic variant wet bench proficiency testing programs by testing for a greater diversity of variants with a wide range of variant allele fractions (VAF) while the validated materials portion is designed to optimize bioinformatics pipelines, augment validations, and assist with pipeline verification after changes to NGS/bioinformatics processes.
• One panel sequencing data file (FASTQ or unaligned BAM), originating from your laboratory and provided to the CAP, for in silico mutagenesis
• Sequencing files containing somatic variants to be downloaded and analyzed by your laboratory bioinformatics pipeline
• Two online activities per year; your CAP shipping contact will be notified via email when the activity is available

Shipping Schedule

• Shipment A: June 17
• Shipment B: December 2