Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens
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 para...
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ftdoajarticles:oai:doaj.org/article:5cd79f0a78a2440c930b202f76e27caf 2023-05-15T15:15:18+02:00 Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum 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 2012-12-01T00:00:00Z https://doi.org/10.1186/1475-2875-11-427 https://doaj.org/article/5cd79f0a78a2440c930b202f76e27caf EN eng BMC http://www.malariajournal.com/content/11/1/427 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-427 1475-2875 https://doaj.org/article/5cd79f0a78a2440c930b202f76e27caf Malaria Journal, Vol 11, Iss 1, p 427 (2012) Multiplex Malaria Antibodies ELISA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2012 ftdoajarticles https://doi.org/10.1186/1475-2875-11-427 2022-12-31T08:20:53Z 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 <0.0001 - 0.004), and antibodies to variants of the same antigen were accurately distinguished within a multiplex reaction. Plasma dilutions of 1:100 or 1:200 were optimal for all antigens for CBA testing. Plasma diluted in a buffer containing 0.05% sodium azide, 0.5% polyvinylalcohol, and 0.8% polyvinylpyrrolidone had the lowest background activity. CBA median fluorescence intensity (MFI) values with 1,000 antigen-conjugated beads/well did not differ significantly from MFI with 5,000 beads/well. CBA and ELISA results correlated well for all antigens except apical membrane antigen-1 (AMA-1). CBA testing produced a greater range of values in samples from malaria endemic areas and less background reactivity for blank samples than ELISA. 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 ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 11 1 |
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ftdoajarticles |
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Multiplex Malaria Antibodies ELISA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Multiplex Malaria Antibodies ELISA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Ondigo Bartholomew N Park Gregory S Gose Severin O Ho Benjamin M Ochola Lyticia A Ayodo George O Ofulla Ayub V John Chandy C Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens |
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Multiplex Malaria Antibodies ELISA Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
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 <0.0001 - 0.004), and antibodies to variants of the same antigen were accurately distinguished within a multiplex reaction. Plasma dilutions of 1:100 or 1:200 were optimal for all antigens for CBA testing. Plasma diluted in a buffer containing 0.05% sodium azide, 0.5% polyvinylalcohol, and 0.8% polyvinylpyrrolidone had the lowest background activity. CBA median fluorescence intensity (MFI) values with 1,000 antigen-conjugated beads/well did not differ significantly from MFI with 5,000 beads/well. CBA and ELISA results correlated well for all antigens except apical membrane antigen-1 (AMA-1). CBA testing produced a greater range of values in samples from malaria endemic areas and less background reactivity for blank samples than ELISA. 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 ... |
format |
Article in Journal/Newspaper |
author |
Ondigo Bartholomew N Park Gregory S Gose Severin O Ho Benjamin M Ochola Lyticia A Ayodo George O Ofulla Ayub V John Chandy C |
author_facet |
Ondigo Bartholomew N Park Gregory S Gose Severin O Ho Benjamin M Ochola Lyticia A Ayodo George O Ofulla Ayub V John Chandy C |
author_sort |
Ondigo Bartholomew N |
title |
Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens |
title_short |
Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens |
title_full |
Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens |
title_fullStr |
Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens |
title_full_unstemmed |
Standardization and validation of a cytometric bead assay to assess antibodies to multiple Plasmodium falciparum recombinant antigens |
title_sort |
standardization and validation of a cytometric bead assay to assess antibodies to multiple plasmodium falciparum recombinant antigens |
publisher |
BMC |
publishDate |
2012 |
url |
https://doi.org/10.1186/1475-2875-11-427 https://doaj.org/article/5cd79f0a78a2440c930b202f76e27caf |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 11, Iss 1, p 427 (2012) |
op_relation |
http://www.malariajournal.com/content/11/1/427 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-427 1475-2875 https://doaj.org/article/5cd79f0a78a2440c930b202f76e27caf |
op_doi |
https://doi.org/10.1186/1475-2875-11-427 |
container_title |
Malaria Journal |
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11 |
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1 |
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1766345661575331840 |