Optimisation and standardisation of a multiplex immunoassay of diverse Plasmodium falciparum antigens to assess changes in malaria transmission using sero-epidemiology.

Wellcome Open Research
Lindsey WuKevin K A Tetteh

Abstract

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 an...Continue Reading

References

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Jun 15, 2018·PLoS Neglected Tropical Diseases·Lou S HermanKevin K A Tetteh

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Citations

Feb 16, 2021·Frontiers in Immunology·Isaac SsewanyanaKevin K A Tetteh
Jun 13, 2021·Nature Communications·Isobel RoutledgeIsabel Rodriguez-Barraquer

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Methods Mentioned

BETA
enzyme-linked immunosorbent assays
ELISAs
in vitro transcription
ELISA

Software Mentioned

cvAUC
R

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