An analytical approach to reduce between-plate variation in multiplex assays that measure antibodies to Plasmodium falciparum antigens

Malaria Journal

Research

Rose F. G. Leke¹  Diane Wallace Taylor²  Naveen K. Bobbili²  John J. Chen³  Andrew Wey³  Rui Fang³ 

[1] Biotechnology Center, Faculty of Medicine and Biomedical Research, University of Yaounde 1, Yaounde, Cameroon;Department of Tropical Medicine, Medical Microbiology and Pharmacology, University of Hawaii John A. Burns School of Medicine, 651 Ilalo Street, BSB 211, 96813, Honolulu, HI, USA;Office of Biostatistics and Quantitative Health Sciences, University of Hawaii John A. Burns School of Medicine, 96813, Honolulu, HI, USA;

关键词: Antibodies;    Between-plate variation;    Multiplex assay;    Normalization procedure;    Malaria;    Placental malaria;   

DOI  :  10.1186/s12936-017-1933-6

 received in 2017-02-13, accepted in 2017-07-11,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundAntibodies 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.MethodsA 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.ResultsCompared 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.ConclusionsFor 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.

【 授权许可】

CC BY   
© The Author(s) 2017

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