title: Simultaneous capture and sequential detection of two malarial biomarkers on magnetic microparticles.
abstract: We have developed a rapid magnetic microparticle-based detection strategy for malarial biomarkers Plasmodium lactate dehydrogenase (pLDH) and Plasmodium falciparum histidine-rich protein II (PfHRPII). In this assay, magnetic particles functionalized with antibodies specific for pLDH and PfHRPII as well as detection antibodies with distinct enzymes for each biomarker are added to parasitized lysed blood samples. Sandwich complexes for pLDH and PfHRPII form on the surface of the magnetic beads, which are washed and sequentially re-suspended in detection enzyme substrate for each antigen. The developed simultaneous capture and sequential detection (SCSD) assay detects both biomarkers in samples as low as 2.0parasites/µl, an order of magnitude below commercially available ELISA kits, has a total incubation time of 35min, and was found to be reproducible between users over time. This assay provides a simple and efficient alternative to traditional 96-well plate ELISAs, which take 5-8h to complete and are limited to one analyte. Further, the modularity of the magnetic bead-based SCSD ELISA format could serve as a platform for application to other diseases for which multi-biomarker detection is advantageous.
authors: Department of Chemistry, Vanderbilt University, Station B 351822, Nashville, TN 37235, USA.
authors: Christine F
authors: CF
authors: Markwalter
authors: Department of Chemistry, Vanderbilt University, Station B 351822, Nashville, TN 37235, USA.
authors: Keersten M
authors: KM
authors: Ricks
authors: Department of Chemistry, Vanderbilt University, Station B 351822, Nashville, TN 37235, USA.
authors: Anna L
authors: AL
authors: Bitting
authors: Department of Chemistry, Vanderbilt University, Station B 351822, Nashville, TN 37235, USA.
authors: Lwiindi
authors: L
authors: Mudenda
authors: Department of Chemistry, Vanderbilt University, Station B 351822, Nashville, TN 37235, USA. Electronic address: david.wright@vanderbilt.edu.
authors: David W
authors: DW
authors: Wright
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000953:Antigens, Protozoan / Q000032:analysis* / Q000097:blood / Q000276:immunology; D015415:Biomarkers / Q000032:analysis / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D018189:Immunomagnetic Separation; D007770:L-Lactate Dehydrogenase / Q000032:analysis* / Q000097:blood / Q000276:immunology; D060328:Magnetic Phenomena; D008288:Malaria; D015800:Protozoan Proteins / Q000032:analysis* / Q000097:blood / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D015415:Biomarkers; C514457:HRP-2 antigen, Plasmodium falciparum; D015800:Protozoan Proteins; D007770:L-Lactate Dehydrogenase
keywords: Immunoassay; Malaria; Plasmodium falciparum histidine-rich protein II; Plasmodium lactate dehydrogenase