Your browser doesn't support javascript.
loading
Validation Strategy for Ultrasensitive Mutation Detection.
Debeljak, Marija; Noë, Michael; Riel, Stacy L; Haley, Lisa M; Norris, Alexis L; Anderson, Derek A; Adams, Emily M; Suenaga, Masaya; Beierl, Katie F; Lin, Ming-Tseh; Goggins, Michael G; Gocke, Christopher D; Eshleman, James R.
Afiliación
  • Debeljak M; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Noë M; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Riel SL; Department of Oncology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Haley LM; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Norris AL; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Anderson DA; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Adams EM; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Suenaga M; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Beierl KF; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Lin MT; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Goggins MG; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Gocke CD; Department of Pathology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
  • Eshleman JR; Department of Oncology, Johns Hopkins University, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
Mol Diagn Ther ; 22(5): 603-611, 2018 10.
Article en En | MEDLINE | ID: mdl-29974360
ABSTRACT

BACKGROUND:

Ultrasensitive detection of low-abundance DNA point mutations is a challenging molecular biology problem, because nearly identical mutant and wild-type molecules exhibit crosstalk. Reliable ultrasensitive point mutation detection will facilitate early detection of cancer and therapeutic monitoring of cancer patients.

OBJECTIVE:

The objective of this study was to develop a method to correct errors in low-level cell line mixes. MATERIALS AND

METHODS:

We tested sample mixes with digital-droplet PCR (ddPCR) and next-generation sequencing.

RESULTS:

We introduced two corrections baseline variant allele frequency (VAF) in the parental cell line was used to correct for copy number variation; and haplotype counting was used to correct errors in cell counting and pipetting. We found ddPCR to have better correlation for detecting low-level mutations without applying any correction (R2 = 0.80) and be more linear after introducing both corrections (R2 = 0.99).

CONCLUSIONS:

The VAF correction was found to be more significant than haplotype correction. It is imperative that various technologies be evaluated against each other and laboratories be provided with defined quality control samples for proficiency testing.
Asunto(s)
Texto completo
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Análisis Mutacional de ADN / Mutación Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Humans Idioma: En Revista: Mol Diagn Ther Asunto de la revista: BIOLOGIA MOLECULAR / FARMACOLOGIA / TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Análisis Mutacional de ADN / Mutación Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Humans Idioma: En Revista: Mol Diagn Ther Asunto de la revista: BIOLOGIA MOLECULAR / FARMACOLOGIA / TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos