新型引物检测 BRAFV600E 突变的敏感性和特异性。
The Sensitivity and Specificity of Novel Primers for Detection of BRAFV600E Mutation.
机构信息
Molecular Pathology and Toxicology, Department of Molecular Microbiology, Central Laboratory in Ibrahim Al-khalil Zakho, Duhok, Kurdistan Region of Iraq.
Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester, UK.
出版信息
Asian Pac J Cancer Prev. 2020 Nov 1;21(11):3191-3198. doi: 10.31557/APJCP.2020.21.11.3191.
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.
目的
本研究旨在评估一种基于 PCR 的新型技术检测癌症早期 BRAF 突变的灵敏度和特异性。
方法
本研究设计并使用了不同长度的引物组，从 8 个碱基到 20 个碱基不等。这些引物是通过应用于癌细胞系开发的。之后，通过进行连续稀释来评估该方法的灵敏度和特异性。
结果
当反应中使用来自细胞系的 150 纳克 DNA 时，长度为 14 个碱基的定量等位基因特异性区分 PCR（QUASAqPCR）引物足以灵敏地检测到 BRAFV600E 与野生型背景的 1:1000 比例（P=0.011）。
结论
QUASA-qPCR 方法的高灵敏度和特异性水平可以提高对癌症早期患者 BRAF 突变检测的诊断准确性，并有助于分层选择适当的治疗方法。
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