Surface plasmon resonance biosensor for rapid label-free detection of microribonucleic acid at subfemtomole level.
Surface plasmon resonance biosensor for rapid label-free detection of microribonucleic acid at subfemtomole level.
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DOI：
10.1021/ac102131s
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发表时间：
2010-12-15
影响因子：
7.4
通讯作者：
Homola, Jiri
中科院分区：
文献类型：
--
作者：
Sipova, Hana;Zhang, Shile;Dudley, Aimee M.;Galas, David;Wang, Kai;Homola, Jiri
MicroRNAs(MiRNAs)与多种调节功能和疾病有关，如癌症和心脏病。因此，它们为未来医疗诊断的检测技术提供了重要的目标。我们报道了一种使用表面等离子体共振(SPR)传感器技术和基于DNA*RNA抗体的分析来快速、灵敏地检测和定量miRNA的新方法。这种方法利用了一种新型的高性能便携式表面等离子体激元传感仪器，该仪器基于一种名为表面等离子体激元耦合器和色散器(SPRCD)的特殊衍射光栅。该光栅的表面被硫代化的DNA寡核苷酸功能化，这种寡核苷酸可以特异性地从液体样品中捕获miRNA，而不需要放大。随后，引入一种识别DNA*RNA杂交物的抗体，以结合到DNA*RNA复合体上，并增强传感器对捕获的miRNA的响应。这种方法允许在不到30分钟的时间内检测到浓度低至下午2点的miRNA，并在高阿托莫尔时检测到绝对量。对小鼠肝组织中miRNA的分析方法进行了评估，发现其结果与定量聚合酶链式反应(QPCR)提供的结果很好地吻合。
MicroRNAs (miRNAs) have been linked with various regulatory functions and disorders, such as cancers and heart diseases. They therefore present an important target for detection technologies for future medical diagnostics. We report here a novel method for rapid and sensitive miRNA detection and quantitation using surface plasmon resonance (SPR) sensor technology and a DNA*RNA antibody-based assay. The approach takes advantage of a novel high-performance portable SPR sensor instrument for spectroscopy of surface plasmons based on a special diffraction grating called a surface plasmon coupler and disperser (SPRCD). The surface of the grating is functionalized with thiolated DNA oligonucleotides which specifically capture miRNA from a liquid sample without amplification. Subsequently, an antibody that recognizes DNA*RNA hybrids is introduced to bind to the DNA*RNA complex and enhance sensor response to the captured miRNA. This approach allows detecting miRNA in less than 30 minutes at concentrations down to 2 pM with an absolute amount at high attomoles. The methodology is evaluated for analysis of miRNA from mouse liver tissues and is found to yield results which agree well with those provided by the quantitative polymerase chain reaction (qPCR).
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