An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens
Abstract Background Antibodies play an important role in immunity to malaria. Recent studies show that antibodies to multiple antigens, as well as, the overall breadth of the response are associated with protection from malaria. Yet, the variability and reliability of antibody measurements against a...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-07-01
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| Series: | Malaria Journal |
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| Online Access: | http://link.springer.com/article/10.1186/s12936-017-1933-6 |
| _version_ | 1818977911138418688 |
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| author | Rui Fang Andrew Wey Naveen K. Bobbili Rose F. G. Leke Diane Wallace Taylor John J. Chen |
| author_facet | Rui Fang Andrew Wey Naveen K. Bobbili Rose F. G. Leke Diane Wallace Taylor John J. Chen |
| author_sort | Rui Fang |
| collection | DOAJ |
| description | Abstract Background Antibodies play an important role in immunity to malaria. Recent studies show that antibodies to multiple antigens, as well as, the overall breadth of the response are associated with protection from malaria. Yet, the variability and reliability of antibody measurements against a combination of malarial antigens using multiplex assays have not been well characterized. Methods A normalization procedure for reducing between-plate variation using replicates of pooled positive and negative controls was investigated. Sixty test samples (30 from malaria-positive and 30 malaria-negative individuals), together with five pooled positive-controls and two pooled negative-controls, were screened for antibody levels to 9 malarial antigens, including merozoite antigens (AMA1, EBA175, MSP1, MSP2, MSP3, MSP11, Pf41), sporozoite CSP, and pregnancy-associated VAR2CSA. The antibody levels were measured in triplicate on each of 3 plates, and the experiments were replicated on two different days by the same technician. The performance of the proposed normalization procedure was evaluated with the pooled controls for the test samples on both the linear and natural-log scales. Results Compared with data on the linear scale, the natural-log transformed data were less skewed and reduced the mean–variance relationship. The proposed normalization procedure using pooled controls on the natural-log scale significantly reduced between-plate variation. Conclusions For malaria-related research that measure antibodies to multiple antigens with multiplex assays, the natural-log transformation is recommended for data analysis and use of the normalization procedure with multiple pooled controls can improve the precision of antibody measurements. |
| first_indexed | 2024-12-20T16:35:15Z |
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| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-12-20T16:35:15Z |
| publishDate | 2017-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-ce69a63ea49f4ebe95113c5ee49c96992022-12-21T19:33:10ZengBMCMalaria Journal1475-28752017-07-0116111010.1186/s12936-017-1933-6An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigensRui Fang0Andrew Wey1Naveen K. Bobbili2Rose F. G. Leke3Diane Wallace Taylor4John J. Chen5Office of Biostatistics and Quantitative Health Sciences, University of Hawaii John A. Burns School of MedicineOffice of Biostatistics and Quantitative Health Sciences, University of Hawaii John A. Burns School of MedicineDepartment of Tropical Medicine, Medical Microbiology and Pharmacology, University of Hawaii John A. Burns School of MedicineBiotechnology Center, Faculty of Medicine and Biomedical Research, University of Yaounde 1Department of Tropical Medicine, Medical Microbiology and Pharmacology, University of Hawaii John A. Burns School of MedicineOffice of Biostatistics and Quantitative Health Sciences, University of Hawaii John A. Burns School of MedicineAbstract Background Antibodies play an important role in immunity to malaria. Recent studies show that antibodies to multiple antigens, as well as, the overall breadth of the response are associated with protection from malaria. Yet, the variability and reliability of antibody measurements against a combination of malarial antigens using multiplex assays have not been well characterized. Methods A normalization procedure for reducing between-plate variation using replicates of pooled positive and negative controls was investigated. Sixty test samples (30 from malaria-positive and 30 malaria-negative individuals), together with five pooled positive-controls and two pooled negative-controls, were screened for antibody levels to 9 malarial antigens, including merozoite antigens (AMA1, EBA175, MSP1, MSP2, MSP3, MSP11, Pf41), sporozoite CSP, and pregnancy-associated VAR2CSA. The antibody levels were measured in triplicate on each of 3 plates, and the experiments were replicated on two different days by the same technician. The performance of the proposed normalization procedure was evaluated with the pooled controls for the test samples on both the linear and natural-log scales. Results Compared with data on the linear scale, the natural-log transformed data were less skewed and reduced the mean–variance relationship. The proposed normalization procedure using pooled controls on the natural-log scale significantly reduced between-plate variation. Conclusions For malaria-related research that measure antibodies to multiple antigens with multiplex assays, the natural-log transformation is recommended for data analysis and use of the normalization procedure with multiple pooled controls can improve the precision of antibody measurements.http://link.springer.com/article/10.1186/s12936-017-1933-6AntibodiesBetween-plate variationMultiplex assayNormalization procedureMalariaPlacental malaria |
| spellingShingle | Rui Fang Andrew Wey Naveen K. Bobbili Rose F. G. Leke Diane Wallace Taylor John J. Chen An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens Malaria Journal Antibodies Between-plate variation Multiplex assay Normalization procedure Malaria Placental malaria |
| title | An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens |
| title_full | An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens |
| title_fullStr | An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens |
| title_full_unstemmed | An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens |
| title_short | An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens |
| title_sort | analytical approach to reduce between plate variation in multiplex assays that measure antibodies to plasmodium falciparum antigens |
| topic | Antibodies Between-plate variation Multiplex assay Normalization procedure Malaria Placental malaria |
| url | http://link.springer.com/article/10.1186/s12936-017-1933-6 |
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