An 8 minute colorimetric paper-based reverse phase vertical flow serum microarray for screening of hyper IgE syndrome.
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| Abstract |
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 μg 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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| Authors | Philippa Reuterswärd, Jesper Gantelius, Helene Andersson Svahn |
| Journal | The Analyst (Analyst) Vol. 140 Issue 21 Pg. 7327-34 (Nov 07 2015) ISSN: 1364-5528 [Electronic] England |
| PMID | 26365343 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't) |
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