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

Biomedical Microdevices
Ngoc M PhamEmmanuel Delamarche

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...Continue Reading

References

Jul 1, 1996·Histochemistry and Cell Biology·P M Lackie
Jan 4, 2003·Analytical Chemistry·David JunckerEmmanuel Delamarche
Dec 21, 2006·Lab on a Chip·Martin ZimmermannEmmanuel Delamarche
Feb 12, 2011·PLoS Medicine·UNKNOWN malERA Consultative Group on Diagnoses and Diagnostics
Sep 14, 2011·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·M L McMorrowS P Kachur
Jun 4, 2014·Journal of Midwifery & Women's Health·Sharon Bernecki DeJoy
Aug 25, 2016·Angewandte Chemie·Gaolian XuJonathan M Cooper
Mar 28, 2017·Malaria Journal·Alfons JimenezXavier C Ding
May 22, 2018·Biomedical Microdevices·Ngoc M PhamEmmanuel Delamarche
Mar 30, 2019·Parasite Epidemiology and Control·Israel CruzJoseph M Ndung'u
Apr 4, 2019·Bulletin of the World Health Organization

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Citations

Oct 12, 2020·Trends in Biotechnology·Séverine Le GacPierre Comizzoli
Jul 31, 2021·Journal of Healthcare Engineering·Yuxing ShiWenhao Zhao
Jul 14, 2021·Journal of Microscopy·Prateek Katare, Sai Siva Gorthi

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