Advancing NGS quality control to enable measurement of actionable mutations in circulating tumor DNA.
Cell Rep Methods
; 1(7): 100106, 2021 11 22.
Article
em En
| MEDLINE
| ID: mdl-35475002
ABSTRACT
The primary objective of the FDA-led Sequencing and Quality Control Phase 2 (SEQC2) project is to develop standard analysis protocols and quality control metrics for use in DNA testing to enhance scientific research and precision medicine. This study reports a targeted next-generation sequencing (NGS) method that will enable more accurate detection of actionable mutations in circulating tumor DNA (ctDNA) clinical specimens. To accomplish this, a synthetic internal standard spike-in was designed for each actionable mutation target, suitable for use in NGS following hybrid capture enrichment and unique molecular index (UMI) or non-UMI library preparation. When mixed with contrived ctDNA reference samples, internal standards enabled calculation of technical error rate, limit of blank, and limit of detection for each variant at each nucleotide position in each sample. True-positive mutations with variant allele fraction too low for detection by current practice were detected with this method, thereby increasing sensitivity.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Tumoral Circulante
Tipo de estudo:
Guideline
Limite:
Humans
Idioma:
En
Revista:
Cell Rep Methods
Ano de publicação:
2021
Tipo de documento:
Article
País de afiliação:
Estados Unidos