Glass-bead-based parallel detection of DNA using composite Raman labels
Glass-bead-based parallel detection of DNA using composite Raman labels
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DOI：
10.1002/smll.200500322
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发表时间：
2006-03-01
期刊：
影响因子：
13.3
通讯作者：
Mirkin, CA
中科院分区：
文献类型：
--
作者：
Jin, RC;Cao, YC;Mirkin, CA
生物分子检测在生物医学研究和疾病诊断中变得越来越重要。[1-8]特别是，微阵列是非常有前途的，因为它们允许一个人进行许多测定同时提供快速读出各种标记策略（荧光团，荧光实体，或纳米粒子）。[1-4]这种阵列已广泛应用于基因组学和蛋白质组学研究。[5]在典型的DNA微阵列形式中，用适当的捕获cDNA（> 200个核苷酸）或合成的寡核苷酸链（25-80个核苷酸）点样玻璃基底。[6]荧光团标记的靶DNA链通过与排列的捕获链杂交而被捕获。由于微阵列中的每个DNA点都被定位编码，因此通常可以通过计算机控制的激光扫描仪系统轻松确定结果。在检测生物分子时，使用点样微阵列的替代方法是随机阵列检测方法。[7-13]这种方法使用单个珠子来代替传统微阵列中的点进行测定。在典型的测定中，制备用于不同但特异性靶标的珠粒批次，然后混合。报告基团用于鉴定珠粒、相应的靶标以及是否与靶核酸序列发生反应。随机阵列形式优于微阵列方法的优点在于，珠粒表现出更快的杂交动力学，制造更容易且更便宜，并且不需要复杂的激光扫描仪系统来获得结果。缺点是失去了微阵列提供的位置编码。因此，需要光谱编码特定测定中涉及的每个靶特异性珠的方法。荧光团通常用作随机阵列策略中的报告基团，[8]然而，由于其相对较宽的发射带和不同染料分子之间的能量转移，可以在多路复用方案中同时检测的荧光染料标记的数量有限。此外，必须执行多个激光扫描事件
Biomolecule detection has become increasingly important in biomedical research and disease diagnosis.[1–8] In particular, microarrays are quite promising because they allow one to carry out many assays simultaneously with rapid readout provided by a variety of labeling strategies (flourophores, chemiluminescent entities, or nanoparticles).[1–4] Such arrays have been widely used in genomics and proteomics research.[5] In a typical DNA microarray format, glass substrates are spotted with appropriate capture cDNA (> 200 nucleotides) or synthesized oligonucleotide strands (25–80 nucleotides).[6] Fluorophore-labeled target DNA strands are captured via hybridization to the arrayed capture strands. Since each DNA spot in the microarray has been positionally encoded, the results can be easily determined, typically, by using a computer-controlled laser scanner system.An alternative to the use of spotted microarrays in detecting biomolecules is the random-array approach to detection.[7–13] This approach uses individual beads for an assay in lieu of a spot in a conventional microarray. In a typical assay, batches of beads for different but specific targets are prepared and then mixed. Reporter groups are used to identify the bead, the corresponding target, and whether or not a reaction with the target nucleic acid sequence has taken place. The advantages of the random array format over the microarray approach are that the beads exhibit faster hybridization kinetics, are easier and less expensive to fabricate, and do not require a sophisticated laser scanner system to obtain the results. The disadvantage is that one loses the positional encoding afforded by a microarray. As such, one needs ways of spectroscopically encoding each of the targetspecific beads involved in a particular assay. Fluorophores are commonly used as reporter groups in the random-array strategy,[8] however, due to their relatively broad emission bands and energy transfer between different dye molecules, the number of fluorescent dye labels that can be simultaneously detected in a multiplexing scheme is limited. In addition, one has to perform multiple laser-scanning events