Brunner C, Hoffmann K, Thiele T, Schedler U, Jehle H, Resch-Genger U
Division Biophotonics, BAM Federal Institute for Materials Research and Testing, Richard-Willstätter-Straße 11, 12489, Berlin, Germany.
Anal Bioanal Chem. 2015 Apr;407(11):3181-91. doi: 10.1007/s00216-014-8450-z. Epub 2015 Jan 24.
Commercial platforms consisting of ready-to-use microarrays printed with target-specific DNA probes, a microarray scanner, and software for data analysis are available for different applications in medical diagnostics and food analysis, detecting, e.g., viral and bacteriological DNA sequences. The transfer of these tools from basic research to routine analysis, their broad acceptance in regulated areas, and their use in medical practice requires suitable calibration tools for regular control of instrument performance in addition to internal assay controls. Here, we present the development of a novel assay-adapted calibration slide for a commercialized DNA-based assay platform, consisting of precisely arranged fluorescent areas of various intensities obtained by incorporating different concentrations of a "green" dye and a "red" dye in a polymer matrix. These dyes present "Cy3" and "Cy5" analogues with improved photostability, chosen based upon their spectroscopic properties closely matching those of common labels for the green and red channel of microarray scanners. This simple tool allows to efficiently and regularly assess and control the performance of the microarray scanner provided with the biochip platform and to compare different scanners. It will be eventually used as fluorescence intensity scale for referencing of assays results and to enhance the overall comparability of diagnostic tests.
由印有靶标特异性DNA探针的即用型微阵列、微阵列扫描仪以及数据分析软件组成的商业平台，可用于医学诊断和食品分析中的不同应用，例如检测病毒和细菌的DNA序列。将这些工具从基础研究转移到常规分析、在受监管领域广泛接受以及在医学实践中使用，除了内部检测对照外，还需要合适的校准工具来定期控制仪器性能。在此，我们展示了一种用于商业化基于DNA的检测平台的新型检测适配校准载玻片的开发，该载玻片由通过在聚合物基质中掺入不同浓度的“绿色”染料和“红色”染料而获得的各种强度的精确排列的荧光区域组成。这些染料呈现具有改进光稳定性的“Cy3”和“Cy5”类似物，根据它们与微阵列扫描仪绿色和红色通道常用标记的光谱特性紧密匹配而选择。这个简单的工具能够高效且定期地评估和控制配备生物芯片平台的微阵列扫描仪的性能，并比较不同的扫描仪。它最终将用作荧光强度标准来参考检测结果，并提高诊断测试的整体可比性。
