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目的建立一种实时荧光定量PCR方法,并用该方法检测乳腺癌患者血浆循环DNA水平。方法用QIAamp血液DNA抽提试剂盒提取61例乳腺癌患者和25例健康妇女血浆循环DNA,用实时荧光定量PCR技术(SYBR Green I染料法)进行定量,结果用设定临界域值时的循环数的值(Ct值)表示,并对定量结果进行ROC曲线分析。结果:乳腺癌患者Ct值(27.71±1.41)显著低于健康对照(30.37±1.32,P<0.001),说明乳腺癌患者血浆循环DNA水平显著高于健康对照组;ROC曲线下面积为0.921。结论应用实时PCR技术检测乳腺癌患者血浆循环DNA水平有可能成为一种无创性乳腺癌分子诊断方法。
Objective To establish a real-time fluorescence quantitative PCR method and use this method to detect plasma circulating DNA level in patients with breast cancer. Methods Plasma DNA was extracted from 61 breast cancer patients and 25 healthy women using QIAamp blood DNA extraction kit and quantified by real-time fluorescence quantitative PCR (SYBR Green I dye method). The results were expressed as cycles Number of values (Ct value) that, and the quantitative results of ROC curve analysis. Results: The Ct value of breast cancer patients (27.71 ± 1.41) was significantly lower than that of healthy controls (30.37 ± 1.32, P <0.001), indicating that circulating plasma DNA levels in breast cancer patients were significantly higher than those in healthy controls. The area under the ROC curve was 0.921. Conclusion Real-time PCR detection of plasma circulating DNA levels in patients with breast cancer may become a molecular diagnostic method of non-invasive breast cancer.