Abstract
Deficiencies of vitamin A and iron affect a significant portion of the world's population, and efforts to characterize patterns of these deficiencies are hampered by a lack of measurement tools appropriate for large-scale population-based surveys. Vitamin A and iron are not easily measured directly, so reliable proxy markers for deficiency status have been identified and adopted. Measurement of inflammatory markers is necessary to interpret vitamin A and iron status markers, because circulating levels are altered by inflammation. We developed a multiplex immunoassay method for simultaneous measurement of five markers relevant to assessing inflammation, vitamin A and iron status: α-1-acid glycoprotein, C-reactive protein, retinol binding protein 4, ferritin and soluble transferrin receptor. Serum and plasma specimens were used to optimize the assay protocol. To evaluate assay performance, plasma from 72 volunteers was assayed using the multiplex technique and compared to conventional immunoassay methods for each of the five markers. Results of the new and conventional assay methods were highly correlated (Pearson Correlations of 0.606 to 0.991, p<.0001). Inter-assay imprecision for the multiplex panel varied from 1% to 8%, and all samples fell within the limits of quantification for all assays at a single dilution. Absolute values given by the multiplex and conventional assays differed, indicating a need for further work to devise a new standard curve. This multiplexed micronutrient immunoassay technique has excellent potential as a cost effective tool for use in large-scale deficiency assessment efforts.
Highlights
Micronutrient (MN) malnutrition is a significant public health problem that affects all segments of society in high, middle and low income countries
In this study we describe the development of a multiplexed MN assessment tool (MMAT) which permits the concurrent measurement of five analytes relevant to vitamin A and iron deficiency surveillance–acid glycoprotein (AGP), Creactive protein (CRP), ferritin, retinol binding protein 4 (RBP) and soluble transferrin receptor (sTfR)– from a single 8 mL volume of serum or plasma
Results from the conventional enzyme-linked immunosorbent assay (ELISA) and the MMAT for the set of 72 heparinized plasma specimens were compared by a variety of methods
Summary
Micronutrient (MN) malnutrition is a significant public health problem that affects all segments of society in high, middle and low income countries. Deficiencies in MNs such as iron, vitamin A (VA), iodine, zinc, and folic acid are associated with adverse health outcomes especially in groups such as pregnant women and children [1,2,3]. Improved MN assessment tools will generate high quality national data on deficiency status that contribute to well-designed control programs [4]. They can contribute to monitoring the impact of the progress of MN programs after implementation
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