数字PCR在肿瘤基因突变检测中的应用

Clinical application of digital PCR for accurate detection of gene mutation in cancer patients

研究肿瘤的有效遗传特性和表观遗传改变需要高灵敏度和特异性的检测工具。然而，无论从灵敏度还是特异性上，传统的检测技术无法满足其所需水平。数字PCR（dPCR）突破传统检测的瓶颈达到稀有突变检测中前所未有的准确性。其通过将一个样本分成百万到千万份，分配到不同反应单元且保证每一反应单元只包含一个目标分子的原理，得到绝对定量的数据。dPCR结合商业化的微流控技术，这种基于微滴原理的dPCR逐渐成为检测癌症患者的重要工具，其应用范围从肿瘤异质性检测到患者体液检测，尤其适用于液体活检分析。

The efficient characterization of genetic and epigenetic alterations in oncology requires highly sensitive and specific high throughput procedures. However, the necessary level of sensitivity and specificity were previously unreachable using conventional testing procedures. By partitioning individual target molecules within separated compartments, digital PCR （dPCR） could allow to overcome such limitations and detect with unprecedented accuracy very rare sequences. In such procedure, the sample is diluted such that each individual compartment will contain no more than one target sequences. The assay provides an absolute value and quantitative data. The recent coupling of dPCR procedure with microfluidic systems in commercial platforms should make droplet based digital PCR an essential tool for the management of patients with cancer. Applications range from the analysis of tumor heterogeneity to the analysis of body effluents. Droplet based digital PCR is also particularly suited for the increasing field of liquid biopsy analysis.