Herr Joshua K, Smith Joshua E, Medley Colin D, Shangguan Dihua, Tan Weihong
Center for Research at the Bio/Nano Interface, Department of Chemistry and Shands Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611, USA.
Anal Chem. 2006 May 1;78(9):2918-24. doi: 10.1021/ac052015r.
We have developed a method for the rapid collection and detection of leukemia cells using a novel two-nanoparticle assay with aptamers as the molecular recognition element. An aptamer sequence was selected using a cell-based SELEX strategy in our laboratory for CCRF-CEM acute leukemia cells that, when applied in this method, allows for specific recognition of the cells from complex mixtures including whole blood samples. Aptamer-modified magnetic nanoparticles were used for target cell extraction, while aptamer-modified fluorescent nanoparticles were simultaneously added for sensitive cell detection. Combining two types of nanoparticles allows for rapid, selective, and sensitive detection not possible by using either particle alone. Fluorescent nanoparticles amplify the signal intensity corresponding to a single aptamer binding event, resulting in improved sensitivity over methods using individual dye-labeled probes. In addition, aptamer-modified magnetic nanoparticles allow for rapid extraction of target cells not possible with other separation methods. Fluorescent imaging and flow cytometry were used for cellular detection to demonstrate the potential application of this method for medical diagnostics.
我们开发了一种使用新型双纳米颗粒检测法快速收集和检测白血病细胞的方法，该方法以适体作为分子识别元件。我们实验室采用基于细胞的SELEX策略，针对CCRF-CEM急性白血病细胞筛选出了一种适体序列，将其应用于该方法时，能够从包括全血样本在内的复杂混合物中特异性识别这些细胞。适体修饰的磁性纳米颗粒用于靶细胞提取，同时添加适体修饰的荧光纳米颗粒用于灵敏的细胞检测。结合使用两种类型的纳米颗粒可实现单独使用任何一种颗粒都无法做到的快速、选择性和灵敏检测。荧光纳米颗粒可放大与单个适体结合事件相对应的信号强度，从而比使用单个染料标记探针的方法具有更高的灵敏度。此外，适体修饰的磁性纳米颗粒能够快速提取靶细胞，这是其他分离方法无法实现的。使用荧光成像和流式细胞术进行细胞检测，以证明该方法在医学诊断中的潜在应用。
