Digital PCR
Digital PCR
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DOI：
10.1073/pnas.96.16.9236
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发表时间：
1999-08-03
影响因子：
11.1
通讯作者：
Kinzler, KW
中科院分区：
文献类型：
--
作者：
Vogelstein, B;Kinzler, KW
预期存在于细胞群体的一小部分中的预定义突变的鉴定对于各种基础研究和临床应用是重要的。在这里，我们描述了一种用于将PCR的指数模拟性质转换为适合于此目的的线性数字信号的方法。通过稀释分离单个分子，并通过PCR单独扩增;然后使用荧光探针分别分析每个产物是否存在突变。该方法的可行性通过检测大肠癌患者粪便中的突变型ras癌基因来证明。该方法提供了DNA样品中变异序列比例的可靠和定量测量。
The identification of predefined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. Here, we describe an approach for transforming the exponential, analog nature of the PCR into a linear, digital signal suitable for this purpose. Single molecules are isolated by dilution and individually amplified by PCR; each product is then analyzed separately for the presence of mutations by using fluorescent probes. The feasibility of the approach is demonstrated through the detection of a mutant ras oncogene in the stool of patients with colorectal cancer. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.