Development of a nine-variant reference material panel to standardize cell-free DNA detection.
Anal Bioanal Chem
; 416(18): 4123-4130, 2024 Jul.
Article
en En
| MEDLINE
| ID: mdl-38782781
ABSTRACT
Detection of specific gene mutations in cell-free DNA (cfDNA) serves as a valuable cancer biomarker and is increasingly being explored as an appealing alternative to tissue-based methods. However, the lack of available reference materials poses challenges in accurately evaluating the performance of different assays. In this study, we present the development of a comprehensive reference material panel for cfDNA detection, encompassing nine hotspot mutations in KRAS/BRAF/EGFR/PIK3CA at three variant allele frequencies (VAFs), ranging from 0.33 to 23.9%. To mimic cfDNA, these reference materials were generated by enzymatically digesting cell-line DNA into approximately 154-bp to 173-bp fragments using a laboratory-developed reaction system. The VAFs for each variation were precisely determined through validated digital PCR assays with high accuracy. Furthermore, the reliability and applicability of this panel were confirmed through two independent NGS assays, yielding concordant results. Collectively, our findings suggest that this novel reference material panel holds great potential for validation, evaluation, and quality control processes associated with liquid biopsy assays.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Estándares de Referencia
/
Proteínas Proto-Oncogénicas B-raf
/
Ácidos Nucleicos Libres de Células
Límite:
Humans
Idioma:
En
Revista:
Anal Bioanal Chem
Año:
2024
Tipo del documento:
Article
País de afiliación:
China