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Biomed Microdevices. 2019 Feb 27;21(1):24. doi: 10.1007/s10544-019-0376-y.

Immuno-gold silver staining assays on capillary-driven microfluidics for the detection of malaria antigens.

Author information

1
ETH Zürich, Mobile Health Systems Lab, Department of Health Sciences and Technology, BAA, Institute of Robotics and Intelligent Systems, Lengghalde 5, CH-8092, Zürich, Switzerland.
2
Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.
3
University of Basel, Petersgraben 1, 4001, Basel, Switzerland.
4
Kantonsspital Aarau AG, Medizinische Genetik, Institut für Labormedizin, Tellstrasse 25, CH-5001, Aarau, Switzerland.
5
IBM Research - Zurich, Säumerstrasse 4, CH-8803, Rüschlikon, Switzerland.
6
IBM Research - Zurich, Säumerstrasse 4, CH-8803, Rüschlikon, Switzerland. emd@zurich.ibm.com.

Abstract

Accurate and affordable rapid diagnostic tests (RDTs) are indispensable but often lacking for many infectious diseases. Specifically, there is a lack of highly sensitive malaria RDTs that can detect low antigen concentration at the onset of infection. Here, we present a strategy to improve the sensitivity of malaria RDTs by using capillary-driven microfluidic chips and combining sandwich immunoassays with electroless silver staining. We used 5 μm fluorescent beads functionalized with capture antibodies (cAbs). These beads are self-assembled by capillary action in recessed "bead lanes", which cross the main flow path of chips microfabricated in Si and SU-8. The binding of analytes to detection antibodies (dAbs) and secondary antibodies (2ndAbs) conjugated to gold nanoparticles (NPs) allows the formation of a silver film on the beads. Such silver film masks the fluorescent core of the bead inversely proportional to the concentration of antigen in a sample. We illustrate this method using the recombinant malaria antigen Plasmodium falciparum histidine-rich-protein 2 (rPfHRP2) spiked in human serum. This antigen was a recombinant HRP2 protein expressed in Escherichia coli, which is also the standard reference material. The limit of detection (LOD) of our immunoassay was found to be less than 6 ng mL-1 of rPfHRP2 within 20 min, which is approaching the desired sensitivity needed in the Target Product Profile (TPP) for malaria elimination settings. The concept presented here is flexible and may also be utilized for implementing fluorescence immunoassays for the parallel detection of biomarkers on capillary-driven microfluidic chips.

KEYWORDS:

Capillary; Fluorescent beads; Malaria infection; Microfluidics; PfHRP2; Silver staining immunoassays

PMID:
30810808
DOI:
10.1007/s10544-019-0376-y
[Indexed for MEDLINE]

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