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Related Concept Videos

The Ras Gene02:38

The Ras Gene

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The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a...
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Related Experiment Video

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Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
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Screening for circulating RAS/RAF mutations by multiplex digital PCR.

Rikke Fredslund Andersen1, Anders Jakobsen2

  • 1Department of Clinical Immunology and Biochemistry, Vejle Hospital, Vejle, Denmark.

Clinica Chimica Acta; International Journal of Clinical Chemistry
|May 17, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a sensitive multiplex digital PCR method for detecting multiple cancer gene mutations in plasma. This liquid biopsy approach shows promise for routine clinical use in cancer management.

Keywords:
Digital PCRMultiplex screeningPlasmactDNA

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Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • Liquid biopsies are gaining interest for cancer management, including treatment selection and recurrence monitoring.
  • Detecting circulating tumor DNA (ctDNA) is challenging due to low levels in early-stage disease or treatment response.
  • Technical hurdles limit the widespread clinical application of ctDNA analysis.

Purpose of the Study:

  • To develop and validate a sensitive multiplex digital PCR method for screening multiple cancer-associated mutations in plasma.
  • To assess the clinical utility of this method in cholangiocarcinoma patients.

Main Methods:

  • A multiplex digital PCR assay was developed to screen for 31 mutations in KRAS, NRAS, BRAF, and PIK3CA genes in plasma.
  • The specificity was determined by the limit of blank (0.006%-0.06%).
  • Mutations detected by multiplex analysis were confirmed and quantified using duplex analyses.

Main Results:

  • The method demonstrated high sensitivity and specificity for detecting mutations in plasma.
  • In cholangiocarcinoma patients, all tumor-detected mutations were also found in plasma samples.
  • Perfect agreement was observed for wild-type status between tumor and plasma, with all other mutations testing negative.

Conclusions:

  • The developed multiplex digital PCR method offers a sensitive and simultaneous analysis of multiple gene mutations from plasma.
  • This approach represents a significant step towards the routine clinical application of liquid biopsies in cancer management.
  • The method's high accuracy in detecting mutations and wild-type status supports its potential for clinical decision-making.