期刊
ANALYST
卷 140, 期 21, 页码 7327-7334出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c5an01013f
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资金
- European Research Council under the European Union's Seventh Framework Programme (FP)/ERC [615458]
- Swedish Research Council
Reverse phase microarrays are useful tools for affinity-based detection in hundreds of samples simultaneously. However, current methods typically require long assay times and fluorescent detection. Here we describe a paper-based Vertical Flow Microarray (VFM) assay as a rapid 8-minute colorimetric alternative for reverse phase microarray analysis. The VFM platform was optimized for detection of IgE with a detection limit of 1.9 mu g mL(-1) in whole serum. Optimized conditions were then used to screen 113 serum samples simultaneously for hyper IgE syndrome (hIgE), a rare primary immunodeficiency characterized by elevated levels of IgE. The same set of samples were then analysed with a conventional planar microarray with fluorescent detection for head-to-head testing. Both assays found elevated levels in three out of four hIgE patient samples, whereas no control samples displayed elevated levels in either method. The comparison experiments showed a good correlation between the two assays, as determined from a linear correlation study (Pearson's r = 0.76). Further, the assay-time reduction and reproducibility (intra assay CV = 12.4 +/- 4.11%) demonstrate the applicability of the VFM platform for high throughput reverse phase screening.
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