Quantitative Bead-Based HRP2 Assay

HRP2 was quantified using a highly sensitive laboratory based quantitative bead suspension array (qSA) based on Luminex technology [17 (link)]. After incubating 2’000 magnetic beads per analyte with blood sample at 1:5 dilution, beads were sequentially incubated with in-house biotinylated antibody α-HRP2 (MBS832975, MyBiSource, San Diego, CL) and streptavidin-PE (42250-1ML, Sigma Aldrich, St. Louis, MO). After a final wash, a minimum of 50 microspheres per analyte were acquired using the Luminex xMAP^® 100/200 analyser (Luminex Corp., Austin, TX). After subtracting background (blank) values, median fluorescent intensities (MFI) were normalized to account for plate to plate variation and quantification was performed against a 5-parameter logistic (5-PL) regression curve consisting of recombinant protein HRP2 type A (890015, Microcoat GmbH, Germany).

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Briand V., Cottrell G., Tuike Ndam N., Martiáñez-Vendrell X., Vianou B., Mama A., Kouwaye B., Houzé S., Bailly J., Gbaguidi E., Sossou D., Massougbodji A., Accrombessi M., Mayor A., Ding X.C, & Fievet N. (2020). Prevalence and clinical impact of malaria infections detected with a highly sensitive HRP2 rapid diagnostic test in Beninese pregnant women. Malaria Journal, 19, 188.

Publication 2020

streptavidin-PE

Antibody Austin Blood Dilution Microspheres Recombinant protein Streptavidin

Corresponding Organization :

Other organizations : Inserm, Université de Bordeaux, Institut de Recherche pour le Développement, Université Paris Cité, Mère et Enfant en Milieu Tropical, Barcelona Institute for Global Health, Universitat de Barcelona, Institut National des Recherches Agricoles du Bénin, Assistance Publique – Hôpitaux de Paris, Hôpital Bichat-Claude-Bernard, Foundation for Innovative New Diagnostics

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Variable analysis

independent variables

Dilution of blood sample (1:5)

dependent variables

Median fluorescent intensities (MFI) of HRP2

control variables

Number of magnetic beads per analyte (2,000)
Biotinylated antibody α-HRP2 (MBS832975, MyBiSource, San Diego, CL)
Streptavidin-PE (42250-1ML, Sigma Aldrich, St. Louis, MO)
Luminex xMAP® 100/200 analyser (Luminex Corp., Austin, TX)
Recombinant protein HRP2 type A (890015, Microcoat GmbH, Germany)

controls

Blank (background) values

Annotations

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