An 8 minute colorimetric paper-based reverse phase vertical flow serum microarray for screening of hyper IgE syndrome

Analyst. 2015 Nov 7;140(21):7327-34. doi: 10.1039/c5an01013f.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Area Under Curve
  • Collodion / chemistry
  • Colorimetry / instrumentation*
  • Colorimetry / methods*
  • Equipment Design
  • Humans
  • Immunoglobulin E / analysis*
  • Job Syndrome / blood*
  • Job Syndrome / diagnosis*
  • Limit of Detection
  • Microarray Analysis
  • Paper
  • Protein Array Analysis / methods
  • Reference Values
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Immunoglobulin E
  • Collodion