Sensitivity, specificity, and the hybridization isotherms of DNA chips
Sensitivity, specificity, and the hybridization isotherms of DNA chips
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DOI：
10.1016/s0006-3495(04)74150-8
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发表时间：
2004-02-01
影响因子：
3.4
通讯作者：
Zhulina, EB
中科院分区：
文献类型：
--
作者：
Halperin, A;Buhot, A;Zhulina, EB
表面和整体的竞争性杂交降低了DNA芯片的敏感度。当不同的靶点可以与同一探针杂交时，就会发生竞争性表面杂交。当靶标可以与溶液中的游离互补链杂交时，竞争性本体杂交就发生了。根据斑点的杂交等温线，定量了竞争性杂交对DNA芯片热力学性能的影响。这些都将杂化的平衡度与主体组成联系起来。杂交等温线以朗缪尔等温线的形式出现，对探针层内的静电相互作用进行了修改。平衡分析的灵敏度与等温线的斜率直接相关。一种更简单的描述是可能的，就c(50)值而言，该值指定对应于表面50%杂化的主体成分。竞争杂交的效果对于DNA芯片结果的定量分析很重要，特别是在用于研究点突变时。
Competitive hybridization, at the surface and in the bulk, lowers the sensitivity of DNA chips. Competitive surface hybridization occurs when different targets can hybridize with the same probe. Competitive bulk hybridization takes place when the targets can hybridize with free complementary chains in the solution. The effects of competitive hybridization on the thermodynamically attainable performance of DNA chips are quantified in terms of the hybridization isotherms of the spots. These relate the equilibrium degree of the hybridization to the bulk composition. The hybridization isotherm emerges as a Langmuir isotherm modified for electrostatic interactions within the probe layer. The sensitivity of the assay in equilibrium is directly related to the slope of the isotherm. A simpler description is possible, in terms of c(50) values specifying the bulk composition corresponding to 50% hybridization at the surface. The effects of competitive hybridization are important for the quantitative analysis of DNA chip results, especially when used to study point mutations.