Molecular Oncology Laboratory, Fundación Para La Investigación del Hospital General Universitario De Valencia, Valencia, Spain.
Mixed Unit TRIAL, (Príncipe Felipe Research Centre & Fundación Para La Investigación Del Hospital General Universitario De Valencia), Valencia, Spain.
Expert Rev Mol Diagn. 2021 Jan;21(1):3-15. doi: 10.1080/14737159.2021.1860759. Epub 2020 Dec 30.
: Precision medicine is already a reality in oncology, since biomarker-driven therapies have clearly improved patient survival. Furthermore, a new, minimally invasive strategy termed 'liquid biopsy' (LB) has revolutionized the field by allowing comprehensive cancer genomic profiling through the analysis of circulating tumor DNA (ctDNA). However, its detection requires extremely sensitive and efficient technologies. A powerful molecular tool based on the principle of 'divide and conquer' has emerged to solve this problem. Thus, digital PCR (dPCR) allows absolute and accurate quantification of target molecules.: In this review we will discuss the fundamentals of dPCR and the most common approaches used for partition of samples and quantification. The advantages and limitations of dPCR will be mentioned in the context of LB in oncology.: In our opinion, dPCR has proven to be one of the most sensitive methods available for LB analysis, albeit some aspects such as its capacity of multiplexing and protocol standardization still require further improvements. Furthermore, the increasing sensitivities and lower costs of next generation sequencing (NGS) methods position dPCR as a confirmatory and complementary technique for NGS results which will likely prove to be very useful for treatment monitoring and assessing minimal residual disease.
精准医学在肿瘤学领域已经成为现实,因为基于生物标志物的治疗方法明显提高了患者的生存率。此外,一种新的、微创的策略,称为“液体活检”(LB),通过分析循环肿瘤 DNA(ctDNA),对癌症的全面基因组分析进行了革命性的改变。然而,其检测需要极其敏感和高效的技术。一种基于“分而治之”原理的强大分子工具已经出现,以解决这个问题。因此,数字 PCR(dPCR)允许对目标分子进行绝对和准确的定量。
在这篇综述中,我们将讨论 dPCR 的基本原理和最常用的样品分割和定量方法。将在肿瘤学 LB 的背景下讨论 dPCR 的优缺点。
在我们看来,dPCR 已被证明是 LB 分析中最敏感的方法之一,尽管其多重检测能力和协议标准化等方面仍需要进一步改进。此外,下一代测序(NGS)方法的灵敏度不断提高,成本不断降低,使得 dPCR 成为 NGS 结果的确认和补充技术,这对于治疗监测和评估微小残留疾病可能非常有用。