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Summary
Metagenomic Next Generation Sequencing (mNGS) promises to revolutionize infectious disease diagnostics. However, the analytical performance of this technology is still not well understood, especially with respect to healthcare-related applications. NIST is developing materials and methods that will help validate these technologies, bringing them from the benchtop to the bedside.
Description
Candidate RM 8376 consists of 20 components with DNA from 19 bacterial strains (nominally 50 ng/uL) and 1 human sample (100 ng/uL) to serve as background. Its purpose is to enable developers and regulators to evaluate the performance metrics of metagnomic next generation sequencing (mNGS) tools. The material will be certified for chromosomal copy number based on droplet digital PCR analysis, providing ground truth abundance for making user-defined, fit-for-purpose samples. Assembled genomes will also be included for informational value. These will allow users to evaluate potential sources of mNGS bias and improve overall measurement confidence.
NIST will publish the Report of Analysis (RoA) upon completion of the stability study.
Genomes for all 19 strains are published on the NCBI website.
Droplet digital PCR (ddPCR) assays with primers and probes are described listed in the RoA.
Major Accomplishments
Genome Assemblies of 19 microbial genomes
Homogeneity testing of 20 components