The Sensitivity and Specificity of Novel Primers for Detection of BRAFV600E Mutation

Document Type : Research Articles


1 Molecular Pathology and Toxicology, Department of Molecular Microbiology, Central Laboratory in Ibrahim Al-khalil Zakho, Duhok, Kurdistan Region of Iraq.

2 Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester, UK.


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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