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Updated: Nov 28, 2025
Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
Published on: October 8, 2015
The Sensitivity and Specificity of Novel Primers for Detection of BRAFV600E Mutation
Bizhar A Tayeb1, Howard J Pringle2
1Molecular Pathology and Toxicology, Department of Molecular Microbiology, Central Laboratory in Ibrahim Al-khalil Zakho, Duhok, Kurdistan Region of Iraq.
Objective:
This study aimed to evaluate the sensitivity and specificity of a PCR-based novel technique for the detection of BRAF mutation in early stages of the cancer.
Methods:
Different lengths of primer sets, ranging from 8 bp to 20 bp, were designed and used in this study. These primers were developed by applying on cancer cell lines. After that, the sensitivity and specificity of the methodology was evaluated by making serial dilutions.
Results:
The quantitative allele specific discrimination PCR (QUASAqPCR) primer with 14 bp length was sensitive enough to detect significantly 1:1,000 ratio of BRAFV600E to wild-type background (P = 0.011), when using 150 nanograms of DNA from cell lines in the reactions.
Conclusion:
High sensitivity and specificity levels of QUASA-qPCR method can improve diagnostic accuracy for BRAF mutation testing in patients at early stages of cancers and help stratify the appropriate choice of treatment.
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