Division of Gene Technology, Science for Life Laboratory, Royal Institute of Technology, Stockholm, Sweden.
PLoS One. 2013 Aug 12;8(8):e70504. doi: 10.1371/journal.pone.0070504. eCollection 2013.
Here we present a novel approach entitled Magnetic Forced Hybridization (MFH) that provides the means for efficient and direct hybridization of target nucleic acids to complementary probes immobilized on a glass surface in less than 15 seconds at ambient temperature. In addition, detection is carried out instantly since the beads become visible on the surface. The concept of MFH was tested for quality control of array manufacturing, and was combined with a multiplex competitive hybridization (MUCH) approach for typing of Human Papilloma Virus (HPV). Magnetic Forced Hybridization of bead-DNA constructs to a surface achieves a significant reduction in diagnostic testing time. In addition, readout of results by visual inspection of the unassisted eye eliminates the need for additional expensive instrumentation. The method uses the same set of beads throughout the whole process of manipulating and washing DNA constructs prior to detection, as in the actual detection step itself.
在这里，我们提出了一种新的方法，称为磁强制杂交（Magnetic Forced Hybridization，MFH），它提供了一种在环境温度下，将目标核酸与固定在玻璃表面上的互补探针进行高效和直接杂交的方法，杂交时间不到 15 秒。此外，由于珠子在表面上变得可见，因此可以立即进行检测。我们测试了 MFH 概念用于微阵列制造的质量控制，并将其与多重竞争杂交（Multiplex Competitive Hybridization，MUCH）方法结合，用于人乳头瘤病毒（HPV）的分型。将珠状 DNA 构建体磁强制杂交到表面上，可以显著缩短诊断测试时间。此外，通过肉眼观察结果进行读取，无需额外昂贵的仪器。该方法在整个过程中都使用相同的一组珠子，包括在检测前操作和洗涤 DNA 构建体，以及在实际检测步骤本身。
