Quality metrics for enhanced performance of an NGS panel using single-vial amplification technology.
Subit BaruaSusan J HsiaoEmily ClancyChristopher FreemanMahesh MansukhaniHelen D FernandesPublished in: Journal of clinical pathology (2022)
The assay was highly specific and sensitive with excellent accuracy, reproducibility and repeatability/precision. Concordant results for identification of SNVs and indels were obtained from specimens with DNA input of 2-3 ng, tissue with 10% neoplastic cellularity and variant allelic frequencies of 2.5%-3%. Over 99% of the target areas are shown to achieve at least 500X coverage when parsed through two bioinformatics pipelines. With over 2000 clinical specimens analysed, the success of the panel for reporting of results is 95.3% CONCLUSIONS: The advanced technology enables accurate identification of clinically relevant variants with uniformity of coverage and an impressive turn-around-time. The overall workflow and cost-effectiveness provide added value.
Keyphrases
- affordable care act
- nucleic acid
- bioinformatics analysis
- high throughput
- copy number
- fine needle aspiration
- circulating tumor
- high resolution
- single molecule
- cell free
- sensitive detection
- emergency department
- fluorescent probe
- electronic health record
- quality improvement
- mass spectrometry
- quantum dots
- circulating tumor cells