College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, PR China.
Biosens Bioelectron. 2011 Aug 15;26(12):4697-701. doi: 10.1016/j.bios.2011.05.017. Epub 2011 May 19.
Single nucleotide polymorphism (SNP) genotyping is attracting extensive attentions owing to its direct connections with human diseases including cancers. Here, we have developed a highly sensitive chemiluminescence biosensor based on circular strand-displacement amplification and the separation by magnetic beads reducing the background signal for point mutation detection at room temperature. This method took advantage of both the T4 DNA ligase recognizing single-base mismatch with high selectivity and the strand-displacement reaction of polymerase to perform signal amplification. The detection limit of this method was 1.3 × 10(-16)M, which showed better sensitivity than that of most of those reported detection methods of SNP. Additionally, the magnetic beads as carrier of immobility was not only to reduce the background signal, but also may have potential apply in high through-put screening of SNP detection in human genome.
单核苷酸多态性 (SNP) 基因分型因其与癌症等人类疾病的直接关联而受到广泛关注。在这里，我们开发了一种基于环形链置换扩增的高灵敏度化学发光生物传感器，并通过磁珠分离降低了室温下用于点突变检测的背景信号。该方法利用 T4 DNA 连接酶具有高选择性识别单碱基错配的特性和聚合酶的链置换反应进行信号放大。该方法的检测限为 1.3×10(-16)M，比大多数已报道的 SNP 检测方法具有更好的灵敏度。此外，作为固定化载体的磁珠不仅可以降低背景信号，而且可能在高通量筛选人类基因组中的 SNP 检测方面具有潜在应用。
