Malaria Journal (Dec 2012)

Standardization and validation of a cytometric bead assay to assess antibodies to multiple <it>Plasmodium falciparum</it> recombinant antigens

  • Ondigo Bartholomew N,
  • Park Gregory S,
  • Gose Severin O,
  • Ho Benjamin M,
  • Ochola Lyticia A,
  • Ayodo George O,
  • Ofulla Ayub V,
  • John Chandy C

DOI
https://doi.org/10.1186/1475-2875-11-427
Journal volume & issue
Vol. 11, no. 1
p. 427

Abstract

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Abstract Background Multiplex cytometric bead assay (CBA) have a number of advantages over ELISA for antibody testing, but little information is available on standardization and validation of antibody CBA to multiple Plasmodium falciparum antigens. The present study was set to determine optimal parameters for multiplex testing of antibodies to P. falciparum antigens, and to compare results of multiplex CBA to ELISA. Methods Antibodies to ten recombinant P. falciparum antigens were measured by CBA and ELISA in samples from 30 individuals from a malaria endemic area of Kenya and compared to known positive and negative control plasma samples. Optimal antigen amounts, monoplex vs multiplex testing, plasma dilution, optimal buffer, number of beads required were assessed for CBA testing, and results from CBA vs. ELISA testing were compared. Results Optimal amounts for CBA antibody testing differed according to antigen. Results for monoplex CBA testing correlated strongly with multiplex testing for all antigens (r = 0.88-0.99, P values from Conclusion With optimization, CBA may be the preferred method of testing for antibodies to P. falciparum antigens, as CBA can test for antibodies to multiple recombinant antigens from a single plasma sample and produces a greater range of values in positive samples and lower background readings for blank samples than ELISA.

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