Department of Chemical and Biomolecular Engineering (BK21+Program), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea; School of Natural Science, Ulsan National Institute of Science and Technology, Ulsan 689-798, Republic of Korea.
Department of Chemical and Biomolecular Engineering (BK21+Program), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Biosens Bioelectron. 2015 Oct 15;72:127-32. doi: 10.1016/j.bios.2015.04.093. Epub 2015 Apr 30.
We herein developed a novel colorimetric polydiacetylene (PDA) sensor for very convenient detection of clinical DNA samples based on the interaction between an intercalator and dsDNA. We modified the terminal carboxyl group of a diacetylene monomer (10,12-pentacosadiynoic acid; PCDA) with the intercalator 9-aminoacridine (9AA) and prepared 9AA-modified PDA liposomes containing PCDA-9AA/PCDA/phospholipid (1,2-dimyristoyl-rac-glycero-3-phosphocholine) at a molar ratio of 1.5:6.5:2.0. The PDA sensor underwent an obvious color transition from blue to red in the presence of dsDNA molecules that were PCR-amplified from genomic DNA due to the insertion of the 9AA head group of PDA into the dsDNA. DNA concentrations as low as 20 nM and relatively small molecules (around 100 base pairs) could be detected by the sensor within 1h without DNA electrophoresis. This novel colorimetric method is simple, does not require any instrument, and is therefore appropriate for POCT or portable molecular diagnostic kit.
我们在此开发了一种新型比色聚二乙炔（PDA）传感器，可通过插入物与双链 DNA 之间的相互作用，非常方便地检测临床 DNA 样本。我们用插入物 9-氨基吖啶（9AA）修饰了二乙炔单体（10,12-二十五碳二炔酸；PCDA）的末端羧基，并制备了含有 PCDA-9AA/PCDA/磷脂（1,2-二肉豆蔻酰-sn-甘油-3-磷酸胆碱）的摩尔比为 1.5:6.5:2.0 的 9AA 修饰 PDA 脂质体。由于 PDA 的 9AA 头基插入双链 DNA 中，PDA 传感器在存在从基因组 DNA 扩增的 dsDNA 分子时会发生明显的颜色从蓝色到红色的转变。在 1 小时内，传感器可以检测到低至 20 nM 的 DNA 浓度和相对较小的分子（约 100 个碱基对），而无需 DNA 电泳。这种新型比色法简单，不需要任何仪器，因此适合 POCT 或便携式分子诊断试剂盒。
