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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Optimisation and standardisation of a multiplex immunoassay of diverse plasmodium falciparum antigens to assess changes in malaria transmission using sero-epidemiology.
Wellcome Open Research, Volume 4, Article 26, Year 2019
Notification
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Description
Background: Antibody responses have been used to characterise transmission and exposure history in malaria-endemic settings for over a decade. Such studies have typically been conducted on well-standardised enzyme-linked immunosorbent assays (ELISAs). However, recently developed quantitative suspension array technologies (qSAT) are now capable of high-throughput and multiplexed screening of up to hundreds of analytes at a time. This study presents a customised protocol for the Luminex MAGPIX© qSAT using a diverse set of malaria antigens. The aim is to develop a standardised assay for routine serological surveillance that is implementable across laboratories and epidemiological settings. Methods: A panel of eight Plasmodium falciparum recombinant antigens, associated with long-and short-lived antibody responses, was designed for the Luminex MAGPIX© platform. The assay was optimised for key steps in the protocol: antigen-bead coupling concentration, buffer composition, serum sample dilution, and bead storage conditions. Quality control procedures and data normalisation methods were developed to address high-throughput assay processing. Antigen-specific limits of quantification (LOQs) were also estimated using both in-house and WHO reference serum as positive controls. Results: Antigen-specific bead coupling was optimised across five serum dilutions and two positive controls, resulting in concentrations operational within stable analytical ranges. Coupled beads were stable after storage at room temperature (22⁰C) for up to eight weeks. High sensitivity and room temperature (22⁰C) for up to eight weeks. High sensitivity and specificity for distinguishing positive and negative controls at serum sample dilutions of 1:500 (AUC 0.94 95%CI 0.91-0.96) and 1:1000 (AUC 0.96 95%CI 0.94-0.98) were observed. LOQs were also successfully estimated for all analytes but varied by antigen and positive control. Conclusions: This study demonstrates that developing a standardised malaria-specific qSAT protocol for a diverse set of antigens is achievable, though further optimisations may be required. Quality control and data standardisation methods may also be useful for future analysis of large sero-epidemiological surveys.
Authors & Co-Authors
Wu, Lindsey
Unknown Affiliation
Hall, Tom
Unknown Affiliation
Ssewanyana, Isaac
Unknown Affiliation
Oulton, Tate
Unknown Affiliation
Patterson, Catriona
Unknown Affiliation
Vasileva, Hristina
Unknown Affiliation
Singh, Susheel Kumar
Unknown Affiliation
Affara, Muna
Unknown Affiliation
Mwesigwa, Julia
Unknown Affiliation
Correa, Simon
Unknown Affiliation
Bah, Mamadou Djoulde
Unknown Affiliation
D'Alessandro, Umberto
Unknown Affiliation
Sepúlveda, Nuno
Unknown Affiliation
Drakeley, Chris J.
Unknown Affiliation
Tetteh, Kevin Kweku Adjei
Unknown Affiliation
Statistics
Citations: 42
Authors: 15
Affiliations: 6
Identifiers
Doi:
10.12688/wellcomeopenres.14950.1
e-ISSN:
2398502X
Research Areas
Infectious Diseases
Study Design
Cross Sectional Study
Study Approach
Quantitative