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. 2024 May 21;24(11):3280.
doi: 10.3390/s24113280.

Point-of-Care Fluorescence Biosensing System for Rapid Multi-Allergen Screening

Silvia Demuru  1 , Hui Chai-Gao  1 , Yevhen Shynkarenko  1 , Nicola Hermann  1 , Patricia-Daiana Boia  1 , Peter Cristofolini  1 , Bradley Petkus  1 , Silvia Generelli  1 , Samantha Paoletti  1 , Stefano Cattaneo  1 , Loïc Burr  1
Affiliations

Point-of-Care Fluorescence Biosensing System for Rapid Multi-Allergen Screening

Silvia Demuru et al. Sensors (Basel). .

Abstract

With the steady increase in allergy prevalence worldwide, there is a strong need for novel diagnostic tools for precise, fast, and less invasive testing methods. Herein, a miniatured fluorescence-based biosensing system is developed for the rapid and quantitative detection of allergen-specific immunoglobulin-E. An antibody-based fluorescence assay in a microfluidic-patterned slide, combined with a custom-made portable fluorescence reader for image acquisition and user-friendly software for the data analysis, enables obtaining results for multiple allergens in just ~1 h with only 80 μL of blood serum. The multiplexed detection of common birch, timothy grass, cat epithelia, house dust mite, and dog epithelia shows quantitative IgE-mediated allergic responses to specific allergens in control serum samples with known total IgE concentration. The responses are verified with different control tests and measurements with a commercial fluorescence reader. These results open the door to point-of-care allergy screening for early diagnosis and broader access and for large-scale research in allergies.

Keywords: IgE; allergen; allergy; antibody; biosensor; fluorescence; microfluidics; optical reader; point of care; portable reader.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Portable system for allergen screening. (a) Image showing the microfluidic slide with a zoom on the micropillars; (b) simplified schematic of the biosensing assay on the slide; (c) image of the portable reader with a slide inserted; (d) the fluorescent signals on the micropillars with an image acquired with the portable reader and custom software.
Figure 2
Figure 2
Portable reader components and comparison. (a) Schematic of the different components inside the developed portable reader; (b) Comparison of the data acquired by InnoScan and the portable reader, analyzing the microfluidic slide with different gradients of the fluorescent dye molecules Atto-BSA. Lower part shows correspondent line profile of the gradient from the lowest to the higher (24 points from about 10 to 5000 dye mol/μm2).
Figure 3
Figure 3
Images showing the custom software features. (a) Portable reader connected to the laptop with the program and login into the user interface; (b) the feature inside the program to acquire the image of the slide; (c) automated pillar recognition; (d) automated fluorescence data extraction and possibility to export in different Excel reports; the colors represent different fluorescence intensities.
Figure 4
Figure 4
Results for specific IgE detection. (a) Optical image acquired by InnoScan and the portable reader, using a microfluidic slide with different recombinant allergens; (b) Relative fluorescence signals extracted and converted in dye molecules/μm2 based on the internal Atto BSA dye calibration. Tested with control serum (274 IU/mL total IgE). The dashed lines indicate the signal over which there is a significant IgE signal, highlighting the maximum cross-sensitivity signal from the IgG negative control.
See this image and copyright information in PMC

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