【摘要】 数字PCR是近年来迅速发展起来的一种定量分析技术。与传统定量PCR技术不同的是数字PCR不依赖于扩增曲线的循环阈值(CT)进行定量,不受扩增效率的影响,也不必采用看家基因和标准曲线,具有很好的准确度和重现性,可以实现绝对定量分析。迄今为止,已有Fluidigm、Bio-Rad等几家公司相继推出了数字PCR产品,这些产品已经在单细胞分析、癌症早期诊断和产前诊断等研究领域显示出巨大的技术优势和应用前景。本文在现有文献基础上,对数字PCR技术的基本原理和定量方法进行介绍,并对该技术的分类及其特点以及目前的主要应用领域进行评述。更多还原

【Abstract】 Digital PCR,a new technique derived from traditional PCR and quantitative PCR,has revealed great potential in genetic profiling,prenatal diagnosis,cancer related allel and mutation detection.Digital PCR usually involves two steps: partitioning of limiting dilution sample into tens to thousands of individual reactions followed by PCR amplification and endpoint detection to identify the presence or absence of template molecules in each reaction.Different with real time qPCR which greatly depends on cycle threshold(CT) and requires house-keeping gene and standard curve for relative and absolute quantification,digital PCR performs quantification through direct counting of the wells with positive amplification or calculating the concentration based on Poisson algorithm.It minimizes the measurement uncertainty caused by variation of amplification efficiency and has higher accuracy and reproducibility.In recent years,great efforts have been made to develop new methods for digital PCR to improve its sensitivity and reduce its cost.More and more techniques were commercialized soon after it was freshly published by large companies including Bio-Rad,Life technology and so on.In this article,the principle of digital PCR and its quantification method,different types of this technique and its practical applications are reviewed and discussed.更多还原