Novel calibration tools and validation concepts for microarray-based platforms used in molecular diagnostics and food safety control
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- 作者：C. Brunner ; K. Hoffmann ; T. Thiele ; U. Schedler…
- 关键词：New reference material ; Microarray ; Fluorescence ; Standard ; Calibration slide
- 刊名：Analytical and Bioanalytical Chemistry
- 出版年：2015
- 出版时间：April 2015
- 年：2015
- 卷：407
- 期：11
- 页码：3181-3191
- 全文大小：2,525 KB
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文摘
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.