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Nutritional Methodology

Combined Measurement of Ferritin, Soluble Transferrin Receptor, Retinol Binding Protein, and C-Reactive Protein by an Inexpensive, Sensitive, and Simple Sandwich Enzyme-Linked Immunosorbent Assay Technique¹

Juergen G. Erhardt^*,2, John E. Estes, Christine M. Pfeiffer^**, Hans K. Biesalski and Neal E. Craft

^* SEAMEO, Regional Center for Community Nutrition, University of Indonesia, Salemba Raya 6, Jakarta 10430, Indonesia; Craft Technologies, Inc., Wilson, NC 27893; ^** National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341; and University of Hohenheim, Institute of Biological Chemistry and Nutrition, 70599 Stuttgart, Germany

²To whom correspondence should be addressed. E-mail: erhardtj{at}uni-hohenheim.de.

The measurement of vitamin A (VA) and iron status is very important in the assessment of nutritional deficiencies. The objective of this research was to develop a sandwich ELISA technique for the simultaneous measurement of ferritin, soluble transferrin receptor, retinol binding protein, and C-reactive protein (CRP) as indicators for VA and iron status. The inclusion of CRP as marker of infection allows for more accurate interpretation of VA and iron status. This is accomplished in a 30-µL serum or plasma sample using an ELISA with different capture and detection antibodies and different dilutions of the sample. Commercially available clinical serum controls were used for calibration purposes. The developed assays were compared to commercially available traditional tests. Regression coefficients comparing both assays were better than 0.84 (P < 0.001). Using a limited sample set, the sandwich ELISA assay produced very similar specificity and sensitivity compared to traditional methods when common cutoff values were applied. Intra- and interassay variability was between 5 and 14% for all tests. The cost of the materials for all 5 measurements decreases to less than $1/sample if a large number of samples is analyzed. Due to the low cost, high throughput, and comparability to traditional tests, this procedure has several advantages for assessing VA and iron status in population surveys.

KEY WORDS: • ELISA • iron deficiency • vitamin A deficiency • low-cost method • infectious status

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