Lee Ju Seok, Song Joon Jin, Deaton Russell, Kim Jin-Woo
Bio/Nano Technology Laboratory, Institute for Nanoscience and Engineering, University of Arkansas, Fayetteville, AR 72701, USA ; Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA ; Graduate Program in Cell and Molecular Biology, University of Arkansas, Fayetteville, AR 72701, USA ; Department of Chemistry, Seoul National University, Seoul 151-747, Republic of Korea.
Biomed Res Int. 2013;2013:310461. doi: 10.1155/2013/310461. Epub 2013 Nov 13.
Microarray is one of the most powerful detection systems with multiplexing and high throughput capability. It has significant potential as a versatile biosensing platform for environmental monitoring, pathogen detection, medical therapeutics, and drug screening to name a few. To date, however, microarray applications are still limited to preliminary screening of genome-scale transcription profiling or gene ontology analysis. Expanding the utility of microarrays as a detection tool for various biological and biomedical applications requires information about performance such as the limits of detection and quantification, which are considered as an essential information to decide the detection sensitivity of sensing devices. Here we present a calibration design that integrates detection limit theory and linear dynamic range to obtain a performance index of microarray detection platform using oligonucleotide arrays as a model system. Two different types of limits of detection and quantification are proposed by the prediction or tolerance interval for two common cyanine fluorescence dyes, Cy3 and Cy5. Besides oligonucleotide, the proposed method can be generalized to other microarray formats with various biomolecules such as complementary DNA, protein, peptide, carbohydrate, tissue, or other small biomolecules. Also, it can be easily applied to other fluorescence dyes for further dye chemistry improvement.
微阵列是最强大的检测系统之一,具有多重检测和高通量能力。作为一个通用的生物传感平台,它在环境监测、病原体检测、医学治疗和药物筛选等诸多领域具有巨大潜力。然而,迄今为止,微阵列的应用仍局限于基因组规模转录谱分析或基因本体分析的初步筛选。要扩大微阵列作为各种生物和生物医学应用检测工具的效用,需要有关检测和定量限等性能的信息,这些信息被视为决定传感设备检测灵敏度的关键信息。在此,我们提出一种校准设计,将检测限理论和线性动态范围相结合,以寡核苷酸阵列作为模型系统来获得微阵列检测平台的性能指标。通过预测或容差区间,针对两种常见的花青荧光染料Cy3和Cy5提出了两种不同类型的检测限和定量限。除了寡核苷酸外,所提出的方法可以推广到其他含有各种生物分子(如互补DNA、蛋白质、肽、碳水化合物、组织或其他小分子生物分子)的微阵列形式。此外,它还可以很容易地应用于其他荧光染料,以进一步改进染料化学。