New detection modality for label-free quantification of DNA in biological samples via superparamagnetic bead aggregation.
New detection modality for label-free quantification of DNA in biological samples via superparamagnetic bead aggregation.
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DOI：
10.1021/ja300839n
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发表时间：
2012-03-28
影响因子：
15
通讯作者：
Landers, James P.
中科院分区：
文献类型：
--
作者：
Leslie, Daniel C.;Li, Jingyi;Strachan, Briony C.;Begley, Matthew R.;Finkler, David;Bazydlo, Lindsay A. L.;Barker, N. Scott;Haverstick, Doris M.;Utz, Marcel;Landers, James P.
在旋转磁场中结合DNA和超顺磁珠产生视觉上引人注目的多颗粒聚集体，并且能够以皮克/微升范围内的水平进行无标记光学检测和DNA定量。生物样品中的DNA可以直接通过简单地分析放置在磁力搅拌器上的微流体威尔斯孔的光学图像来定量，而无需DNA纯化。聚合由DNA/珠相互作用引起，所述DNA/珠相互作用由离液剂（聚合的非特异性触发剂）的存在或与功能化珠上的寡核苷酸杂交（序列特异性）驱动。本文展示了DNA的定量与灵敏度相媲美的最好的目前可用的荧光测定。该方法的稳健性和灵敏度使其能够广泛应用，在此通过计数真核细胞来说明。该方法使用广泛可用且廉价的台式硬件，为更昂贵的DNA检测和细胞计数技术提供了一种高度可及的低技术微型替代方案。
Combining DNA and superparamagnetic beads in a rotating magnetic field produces multiparticle aggregates that are visually striking, and enables label-free optical detection and quantification of DNA at levels in the picogram per microliter range. DNA in biological samples can be quantified directly by simple analysis of optical images of microfluidic wells placed on a magnetic stirrer without DNA purification. Aggregation results from DNA/bead interactions driven either by the presence of a chaotrope (a nonspecific trigger for aggregation) or by hybridization with oligonucleotides on functionalized beads (sequence-specific). This paper demonstrates quantification of DNA with sensitivity comparable to that of the best currently available fluorometric assays. The robustness and sensitivity of the method enable a wide range of applications, illustrated here by counting eukaryotic cells. Using widely available and inexpensive benchtop hardware, the approach provides a highly accessible low-tech microscale alternative to more expensive DNA detection and cell counting techniques.
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