Nanoparticle-Based Histidine-Rich Protein-2 Assay for the Detection of the Malaria Parasite Plasmodium falciparum

A nanoparticle-based assay for detection and quantification of Plasmodium falciparum histidine-rich protein 2 (HRP2) in urine and serum is reported. The assay uses magnetic beads conjugated with anti-HRP2 antibody for protein capture and concentration, and antibody-conjugated quantum dots for optica...

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Bibliographic Details
Published in:The American journal of tropical medicine and hygiene 2016-08, Vol.95 (2), p.354-357
Main Authors: Castro-Sesquen, Yagahira E, Kim, Chloe, Gilman, Robert H, Sullivan, David J, Searson, Peter C
Format: Article
Language:English
Subjects:
Antibodies, Protozoan - chemistry
Antigens, Protozoan - urine
Biological Assay
Blotting, Western
Calibration
Gene Expression
Humans
Magnets
Malaria, Falciparum - diagnosis
Malaria, Falciparum - parasitology
Malaria, Falciparum - urine
Nanoparticles - chemistry
Plasmodium falciparum
Plasmodium falciparum - genetics
Plasmodium falciparum - isolation & purification
Protozoan Proteins - urine
Quantum Dots - chemistry
Sensitivity and Specificity
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Description
Summary:A nanoparticle-based assay for detection and quantification of Plasmodium falciparum histidine-rich protein 2 (HRP2) in urine and serum is reported. The assay uses magnetic beads conjugated with anti-HRP2 antibody for protein capture and concentration, and antibody-conjugated quantum dots for optical detection. Western blot analysis demonstrated that magnetic beads allow the concentration of HRP2 protein in urine by 20-fold. The concentration effect was achieved because large volume of urine can be incubated with beads, and magnetic separation can be easily performed in minutes to isolate beads containing HRP2 protein. Magnetic beads and quantum dots conjugated to anti-HRP2 antibodies allows the detection of low concentrations of HRP2 protein (0.5 ng/mL), and quantification in the range of 33-2,000 ng/mL corresponding to the range associated with non-severe to severe malaria. This assay can be easily adapted to a noninvasive point-of-care test for classification of severe malaria.
ISSN:0002-9637
1476-1645
DOI:10.4269/ajtmh.15-0772