A microfluidic chip‐based fluorescent biosensor for the sensitive and specific detection of label‐free single‐base mismatch via magnetic beads‐based “sandwich” hybridization strategy
A microfluidic chip‐based fluorescent biosensor for the sensitive and specific detection of label‐free single‐base mismatch via magnetic beads‐based “sandwich” hybridization strategy
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DOI：
10.1002/elps.201300131
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发表时间：
2013-08
期刊：
影响因子：
2.9
通讯作者：
Zongwen Wang;Yingwei Fan;Jinfa Chen;Ying Guo;Weihua Wu;Ye He;Liangjun Xu;Fengfu Fu
中科院分区：
文献类型：
--
作者：
Zongwen Wang;Yingwei Fan;Jinfa Chen;Ying Guo;Weihua Wu;Ye He;Liangjun Xu;Fengfu Fu
本研究开发了一种新型的微流控芯片荧光DNA生物传感器，该传感器利用电泳法驱动模式和磁珠为基础的“三明治”杂交策略，用于单碱基错配DNA的灵敏和超特异性检测。与以往的生物传感器相比，该生物传感器对真实唾液和血清样本的复杂基质具有更强的抵抗力，分析时间更短，对单碱基错配的检测具有更高的分辨能力。这些特点以及它的制造简单、操作方便、稳定性好、可重用性好、成本低等特点，使其成为临床诊断中SNPs基因分型/检测的一种有前途的替代方案。利用该生物传感器成功地检测了唾液和血清中的口腔癌相关基因，无需样品标记和任何预分离或稀释，检出限为5.6×10−11M，相对标准偏差为5%，一碱基错配的判别因子为3.5 8~4.5 4。
A novel microfluidic chip‐based fluorescent DNA biosensor, which utilized the electrophoretic driving mode and magnetic beads‐based “sandwich” hybridization strategy, was developed for the sensitive and ultra‐specific detection of single‐base mismatch DNA in this study. In comparison with previous biosensors, the proposed DNA biosensor has much more robust resistibility to the complex matrix of real saliva and serum samples, shorter analysis time, and much higher discrimination ability for the detection of single‐base mismatch. These features, as well as its easiness of fabrication, operation convenience, stability, better reusability, and low cost, make it a promising alternative to the SNPs genotyping/detection in clinical diagnosis. By using the biosensor, we have successfully determined oral cancer‐related DNA in saliva and serum samples without sample labeling and any preseparation or dilution with a detection limit of 5.6 × 10−11 M, a RSD (n = 5) < 5% and a discrimination factor of 3.58–4.54 for one‐base mismatch.