School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
Nanoscale. 2013 Jun 7;5(11):5027-35. doi: 10.1039/c3nr01010d. Epub 2013 May 2.
We report herein the development of a highly sensitive and selective approach for label-free DNA detection by combining target-recycled ligation (TRL), magnetic nanoparticle assisted target capture/separation, and efficient enzymatic amplification. We show that our approach can detect as little as 30 amol (600 fM in 50 μL) of unlabelled single-stranded DNA targets and offer an exquisitely high discrimination ratio (up to >380 fold with background correction) between a perfect-match cancer mutant and its single-base mismatch (wild-type) DNA target. Furthermore, it can quantitate the rare cancer mutant (KRAS codon 12) in a large excess of coexisting wild-type DNAs down to 0.75%. This sensor appears to be well-suited for sensitive SNP detection and a wide range of DNA mutation based diagnostic applications.
我们在此报告了一种通过结合目标循环连接（TRL）、磁性纳米粒子辅助目标捕获/分离和高效酶扩增，实现无标记 DNA 检测的高灵敏度和选择性方法。我们表明，我们的方法可以检测低至 30 飞摩尔（600 皮摩尔在 50 μL 中）的未标记单链 DNA 靶标，并在完美匹配的癌症突变体与其单碱基错配（野生型）DNA 靶标之间提供极高的区分率（背景校正后高达 >380 倍）。此外，它可以定量检测大量共存的野生型 DNA 中罕见的癌症突变体（KRAS 密码子 12），低至 0.75%。这种传感器似乎非常适合用于敏感 SNP 检测和广泛的基于 DNA 突变的诊断应用。
