Kinetic fingerprinting to identify and count single nucleic acids.
Kinetic fingerprinting to identify and count single nucleic acids.
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DOI：
10.1038/nbt.3246
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发表时间：
2015-07
影响因子：
46.9
通讯作者：
Walter NG
中科院分区：
文献类型：
--
作者：
Johnson-Buck A;Su X;Giraldez MD;Zhao M;Tewari M;Walter NG
MicroRNAs(MiRNAs)已成为一种很有前途的诊断生物标志物。我们介绍了一种称为通过平衡泊松采样的单分子识别(SiMREPS)的动态指纹分析方法，用于单个未标记miRNA分子的无扩增计数，它绕过了特异性的热力学限制，几乎消除了假阳性。我们展示了在缓冲液和最低限度处理的生物液体中对合成和内源性miRNAs的高置信度单分子检测，以及单核苷酸多态之间的500倍区分。
MicroRNAs (miRNAs) have emerged as promising diagnostic biomarkers. We introduce a kinetic fingerprinting approach called Single Molecule Recognition through Equilibrium Poisson Sampling (SiMREPS) for the amplification-free counting of single unlabeled miRNA molecules, which circumvents thermodynamic limits of specificity and virtually eliminates false positives. We demonstrate high-confidence single-molecule detection of synthetic and endogenous miRNAs in both buffer and minimally treated biological liquids, as well as >500-fold discrimination between single nucleotide polymorphisms.