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首页> 外文期刊>Mycological Research >Use of hybridization melting kinetics for detecting Phytophthora species using three-dimensional microarrays: demonstration of a novel concept for the differentiation of detection targets
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Use of hybridization melting kinetics for detecting Phytophthora species using three-dimensional microarrays: demonstration of a novel concept for the differentiation of detection targets

机译：杂交融解动力学在三维立体阵列中检测疫霉菌的应用：展示区分检测目标的新概念

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摘要

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.
机译：基于微阵列的检测受到每种阵列中所用探针多样性的可变和不一致的杂交强度的限制。在本文中，我们介绍了一种利用双链熔解动力学区分检测目标的新概念。在PamChip微阵列上开发了一种微阵列测定法，可通过使用探针-靶标双链体的解链动力学来区分靶疫霉菌。在大多数情况下，靶标和非靶标双链体的杂交动力学显着不同。发现由具有靶标和非靶标DNA的探针形成的双链体的解链动力学分析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可靠，并且能够检测单个疫霉菌种及其混合物。

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