Quantitative PCR–Based Method to Assess Cell-Free DNA Quality, Adjust Input Mass, and Improve Next-Generation Sequencing Assay Performance

Autor:	Seng Lor Saelee, Alexander F. Lovejoy, Bernd Hinzmann, Katrina Mayol, Samantha Huynh, Amy Harrell, Josh Lefkowitz, Niharika Deodhar, Gladys Garcia-Montoya, Stephanie J. Yaung, Daniel M. Klass
Rok vydání:	2022
Předmět:	Mutation High-Throughput Nucleotide Sequencing Humans Molecular Medicine Cell-Free Nucleic Acids Polymerase Chain Reaction Circulating Tumor DNA Pathology and Forensic Medicine
Zdroj:	The Journal of Molecular Diagnostics. 24:566-575
ISSN:	1525-1578
DOI:	10.1016/j.jmoldx.2022.02.005
Popis:	Cell-free (cf)DNA-based testing has undergone increasingly wide adoption, including assays for the detection of circulating tumor DNA. Due to nucleosome protection, cfDNA has a distinctive fragment size of 160 to 180 bp. However, cfDNA can be contaminated with high molecular weight genomic DNA from blood cells released in plasma during sample collection. Such contamination can lead to decreased sensitivity or inconsistent results in cfDNA next-generation sequencing assays. This article describes a technical advancement in which a quantitative PCR method is used for high molecular weight contamination assessment and input mass adjustment, and has been demonstrated to improve consistency of performance in a circulating tumor DNA next-generation sequencing workflow.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f45f83a9e84d89629d0efe4c1a262850 https://doi.org/10.1016/j.jmoldx.2022.02.005 Zobrazit plný text záznamu