State Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, College of Science, Huazhong Agricultural University, Wuhan 430070, PR China.
State Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, College of Science, Huazhong Agricultural University, Wuhan 430070, PR China.
Biosens Bioelectron. 2017 Jun 15;92:321-327. doi: 10.1016/j.bios.2016.11.005. Epub 2016 Nov 5.
Highly sensitive and selective detection of specific DNA sequences is of great importance in clinical diagnosis, environmental and food monitoring, but it still remains challenges to develop a facile method for real sample detection in aqueous solution. Here, a simple and recyclable surface enhanced Raman scattering (SERS) sensor was constructed for Bacillus thuringiensis (Bt) special gene fragment detection by FeO magnetic beads (MBs) and Au-Ag core-shell nanorods (Au@Ag NRs). A hairpin DNA with sulfhydryl and biotin was attached to Au@Ag NRs as indicator, and MBs with streptavidin (SA) were acted as the capture probe. On the basis of the biotin-SA specific interaction, target sequences were first hybridized with the hairpin DNA and exposed the biotin. Subsequently, the Au@Ag NRs were captured by the streptavidin modified MBs, which reduced the suspended NRs and led to the change of Raman intensity. Under the optimal conditions, the SERS intensity revealed a good linearity with Bt transgene fragment ranging from 0.1pM to 1nM with a detection limit of 0.14pM (S/N=3). To demonstrate the specificity of the strategy, the single-base mismatch in DNA was discussed in the SERS assay. The results showed that the sensitivity and accuracy of the proposed method was acceptable in DNA detection, revealing a great potential in special gene detection.
高灵敏度和选择性地检测特定的 DNA 序列在临床诊断、环境和食品监测中非常重要,但开发用于在水溶液中进行实际样品检测的简便方法仍然具有挑战性。在这里,通过 FeO 磁性珠 (MBs) 和 Au-Ag 核壳纳米棒 (Au@Ag NRs) 构建了一种简单且可回收的表面增强拉曼散射 (SERS) 传感器,用于检测苏云金芽孢杆菌 (Bt) 特殊基因片段。具有巯基和生物素的发夹 DNA 被连接到 Au@Ag NRs 上作为指示剂,而带有链霉亲和素 (SA) 的 MBs 则作为捕获探针。基于生物素-SA 的特异性相互作用,首先将目标序列与发夹 DNA 杂交,并暴露出生物素。随后,Au@Ag NRs 被链霉亲和素修饰的 MBs 捕获,这减少了悬浮的 NRs,导致拉曼强度发生变化。在最佳条件下,SERS 强度与 Bt 转基因片段呈现出良好的线性关系,其线性范围为 0.1pM 至 1nM,检测限为 0.14pM (S/N=3)。为了证明该策略的特异性,在 SERS 测定中讨论了 DNA 中的单碱基错配。结果表明,该方法在 DNA 检测中的灵敏度和准确性是可以接受的,这表明它在特殊基因检测方面具有很大的潜力。