Anderson Nari, Szemes Marianna, O'Brien Phil, de Weerdt Marjanne, Schoen Cor, Boender Piet, Bonants Peter
Murdoch University, South Street, Murdoch 6150, Perth Australia.
Mycol Res. 2006 Jun;110(Pt 6):664-71. doi: 10.1016/j.mycres.2006.04.003. Epub 2006 Jun 12.
Microarray-based detection is limited by variable and inconsistent hybridization intensities across the diversity of probes used in each array. In this paper, we introduce a novel concept for the differentiation of detection targets using duplex melting kinetics. A microarray assay was developed on a PamChip microarray enabling the differentiation of target Phytophthora species using the melting kinetics of probe-target duplexes. In the majority of cases the hybridization kinetics of target and non-target duplexes differed significantly. Analysis of the melting kinetics of duplexes formed by probes with target and non-target DNA was found to be an effective method for determining specific hybridization and was independent of fluctuations in hybridization signal intensity. This form of analysis was more robust than the traditional approach based on hybridization intensity, and enabled the detection of individual Phytophthora species and mixtures thereof.
基于微阵列的检测受到每个阵列中使用的各种探针杂交强度可变且不一致的限制。在本文中，我们引入了一种利用双链体解链动力学区分检测靶标的新概念。在PamChip微阵列上开发了一种微阵列分析方法，能够利用探针 - 靶标双链体的解链动力学区分疫霉属目标物种。在大多数情况下，靶标和非靶标双链体的杂交动力学有显著差异。发现分析由探针与靶标和非靶标DNA形成的双链体的解链动力学是确定特异性杂交的有效方法，并且与杂交信号强度的波动无关。这种分析形式比基于杂交强度的传统方法更稳健，并且能够检测单个疫霉属物种及其混合物。
