title: Immunomagnetic capture and colorimetric detection of malarial biomarker Plasmodium falciparum lactate dehydrogenase.
abstract: We report a sensitive, magnetic bead-based colorimetric assay for Plasmodium falciparum lactate dehydrogenase (PfLDH) in which the biomarker is extracted from parasitized whole blood and purified based on antigen binding to antibody-functionalized magnetic particles. Antigen-bound particles are washed, and PfLDH activity is measured on-bead using an optimized colorimetric enzyme reaction (limit of detection [LOD] = 21.1 ± 0.4 parasites/μl). Enhanced analytical sensitivity is achieved by removal of PfLDH from the sample matrix before detection and elimination of nonspecific reductases and species that interfere with the optimal detection wavelength for measuring assay development. The optimized assay represents a simple and effective diagnostic strategy for P. falciparum malaria with time-to-result of 45 min and detection limits similar to those of commercial enzyme-linked immunosorbent assay (ELISA) kits, which can take 4-6 h. This method could be expanded to detect all species of malaria by switching the capture antibody on the magnetic particles to a pan-specific Plasmodium LDH antibody.
authors.affiliation: Department of Chemistry, Vanderbilt University, Nashville, TN 37235, USA.
authors.forename: Christine F
authors.initials: CF
authors.lastname: Markwalter
authors.affiliation: Department of Chemistry, Vanderbilt University, Nashville, TN 37235, USA. Electronic address: k.davis@vanderbilt.edu.
authors.forename: Keersten M
authors.initials: KM
authors.lastname: Davis
authors.affiliation: Department of Chemistry, Vanderbilt University, Nashville, TN 37235, USA. Electronic address: david.wright@vanderbilt.edu.
authors.forename: David W
authors.initials: DW
authors.lastname: Wright
mesh_terms: D003124:Colorimetry / Q000191:economics / Q000379:methods; D057075:Enzyme Assays / Q000191:economics / Q000379:methods; D006801:Humans; D018189:Immunomagnetic Separation / Q000191:economics / Q000379:methods*; D007770:L-Lactate Dehydrogenase / Q000032:analysis* / Q000302:isolation & purification; D057230:Limit of Detection; D016778:Malaria, Falciparum / Q000097:blood / Q000175:diagnosis* / Q000469:parasitology; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000302:isolation & purification*
chemical_list: D007770:L-Lactate Dehydrogenase
keywords: Activity assay; Immunomagnetic capture; Malaria; Plasmodium lactate dehydrogenase