Abstract
BackgroundSerological markers are potentially useful tools for monitoring the progress of malaria control programs, but a better understanding of antibody response dynamics is necessary. The use of a magnetic bead-based immunoassay (MBA) is advantageous compared to ELISA, due to its multiplexing capacity, but limited information is available on the standardization and validation of this assay.MethodsSeveral parameters for multiplex testing of antibodies to Plasmodium antigens were analysed using a set of 4 antigens and 98 sera from Senegalese rural asymptomatic and urban symptomatic individuals. The 4 antigens included Plasmodium falciparum CSP and PfAMA1 peptides, recombinant P. falciparum MSP4p20 and a Plasmodium malariae CSP (PmCSP) peptide. Comparisons with ELISA were done using MSP4p20 and whole schizont extract (SE) antigens.ResultsThe use of fewer beads (1000 beads per well instead of 2000) and 5 µg of antigen per 106 bead were validated as lower amounts. The use of a carrier protein (BSA) was shown to be critical when using peptides and the effect of a 24 h delayed measures was evaluated (5–25% signal decrease). Analysis of Ab responses showed almost equally high levels and prevalence in all transmission settings. Clear distinctions between rural and urban malaria were noted using PmCSP and SE antigens.ConclusionsThis study underlines the importance of further optimization of the MBA technique and highlights the interest of using multistage/multispecies antigens for surveillance of malaria in endemic settings.
Highlights
Serological markers are potentially useful tools for monitoring the progress of malaria control pro‐ grams, but a better understanding of antibody response dynamics is necessary
Antibody responses occur against a broad array of antigens [7] correlating with susceptibility to clinical and severe malaria [6]
Optimization of multiplex protocols The first aim of this study was the technical optimization of the protocol, including parameters such as the optimal number of beads per well, the importance of BSA-conjugated peptides and proteins, impact of delayed reading on fluorescence intensity
Summary
Serological markers are potentially useful tools for monitoring the progress of malaria control pro‐ grams, but a better understanding of antibody response dynamics is necessary. Ongoing efforts to achieve sustainable field results require strategies for accurate evaluation of malaria exposure to monitor the effectiveness of anti-malaria control measures. For this purpose, measurement of antibody responses to relevant immune markers can be used to evaluate exposure and/or immunity in exposed as well as in naïve populations [4, 5]. The immune control of P. falciparum infection is complex, and requires the combined action of antibodies and cell mediated immune responses against both pre-erythrocytic and blood stages [6]. Antibody responses occur against a broad array of antigens [7] correlating with susceptibility to clinical and severe malaria [6]
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