Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, HuBei 430074, PR China.
Biosens Bioelectron. 2011 Jan 15;26(5):2317-22. doi: 10.1016/j.bios.2010.10.002. Epub 2010 Nov 5.
Here a novel capillary electrophoresis (CE) for simultaneous detection of dual single-base mutations using quantum dot-molecular beacon (QD-MB) probe is described. Two QD-MB probes were designed using 585 and 650-nm emitting CdTe QDs which were covalently conjugated to MBs with different DNA oligonucleotide sequences by amide linkage and streptavidin-biotin binding, respectively. The hybridizations of QD-MB probes with different DNA targets were then monitored by CE, and results indicated that the two QD-MB probes specifically hybridized with their complementary DNA sequences, respectively. Target DNA identification was observed to have a high sensitivity of 16.2 pg in CE. Furthermore, the simultaneous detection of dual single-base mutations in a given DNA oligonucleotide was successfully achieved in CE using above two QD-MB probes. This novel CE-assisted QD-MB biosensor offers a promising approach for simultaneous detection of multiple single-base mutations, and exhibits potential capability in the single nucleotide polymorphism (SNP) analysis and high-sensitivity DNA detection.
本文描述了一种新的毛细管电泳（CE）技术，用于使用量子点-分子信标（QD-MB）探针同时检测双重单碱基突变。使用发射波长为 585nm 和 650nm 的 CdTe QD 设计了两个 QD-MB 探针，它们通过酰胺键共价连接到具有不同 DNA 寡核苷酸序列的 MB 上，并通过链霉亲和素-生物素结合。然后通过 CE 监测 QD-MB 探针与不同 DNA 靶标的杂交，结果表明两个 QD-MB 探针分别与它们的互补 DNA 序列特异性杂交。在 CE 中，目标 DNA 的识别具有 16.2pg 的高灵敏度。此外，使用上述两个 QD-MB 探针在 CE 中成功实现了给定 DNA 寡核苷酸中双重单碱基突变的同时检测。这种新型的 CE 辅助 QD-MB 生物传感器为同时检测多个单碱基突变提供了一种有前途的方法，并在单核苷酸多态性（SNP）分析和高灵敏度 DNA 检测方面表现出潜在的能力。
