Viral diagnostics in the era of digital polymerase chain reaction.
Viral diagnostics in the era of digital polymerase chain reaction.
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DOI：
10.1016/j.diagmicrobio.2012.10.009
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发表时间：
2013-01
影响因子：
2.9
通讯作者：
Jerome, Keith R.
中科院分区：
文献类型：
--
作者：
Sedlak, Ruth Hall;Jerome, Keith R.
与定量聚合酶链式反应(QPCR)不同，数字聚合酶链式反应(DPCR)无需标准曲线即可实现对DNA样本的灵敏和准确的绝对定量。一个单独的PCR反应被分成许多单独的反应，每个反应都有积极或消极的信号。通过应用泊松统计，直接根据阳性反应和阴性反应的数量计算出原始样本中的DNA分子数量。随着多种商业化的dPCR平台的问世，人们对低病毒载量检测和低丰度突变检测等临床诊断应用越来越感兴趣，其中dPCR可能优于传统的qPCR。我们回顾了当前的文献，证明了dPCR在病毒诊断中的潜在用途，特别是通过绝对量化目标DNA序列和罕见的突变等位基因检测。
Unlike quantitative PCR (qPCR), digital PCR (dPCR) achieves sensitive and accurate absolute quantitation of a DNA sample without the need for a standard curve. A single PCR reaction is divided into many separate reactions that each have a positive or negative signal. By applying Poisson statistics, the number of DNA molecules in the original sample is directly calculated from the number of positive and negative reactions. The recent availability of multiple commercial dPCR platforms has led to increased interest in clinical diagnostic applications, such as low viral load detection and low abundance mutant detection, where dPCR could be superior to traditional qPCR.Here we review current literature that demonstrates dPCR’s potential utility in viral diagnostics, particularly through absolute quantification of target DNA sequences and rare mutant allele detection.
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