Development and Validation of a New Low-Cost Enzyme-Linked Immunoassay for Serum and Dried Blood Spot Thyroglobulin

摘要

Background: Thyroglobulin (Tg), a biomarker of iodine nutrition, can be measured on dried blood spots (DBS), which simplifies collection and transport in surveys. The World Health Organization recommends DBS-Tg for monitoring iodine status in children. It could also be a useful iodine biomarker during pregnancy. However, the Tg antibody (Ab) used in earlier DBS-Tg assays is no longer commercially available. The aims of the present study were: (i) to develop a new low-cost serum and DBS-Tg sandwich enzyme-linked immunosorbent assay for assessment of Tg in population studies; (ii) to check the stability of DBS-Tg during long-term storage; and (iii) to assess within-subject variability in DBS-Tg. Methods: Serum and DBS samples were measured from healthy pregnant women (n=424) with the new assays, as well as the Immulite 2000 (Siemens), including TgAb positive (n=150) and TgAb negative (n=274) women. DBS-Tg stability was tested over 15 weeks of storage at -20 degrees C. Within-subject variability was evaluated over four weeks in four healthy adults. Results: Intra-assay and interassay variability was 4.4-7.3% and 10.1-12.9% for the new serum Tg assay, and 7.6-12.3% and 7.6-16.5% for the DBS-Tg assay. Correlation between the two serum methods was high (r=0.68, p<0.01). Assay performance in all women and those TgAb negative was comparable. Correlation between the new serum Tg assay and the DBS-Tg assay was high (r=0.78, p<0.01), and agreement expressed as a function of the average Tg concentration for the two methods (X) was 0.59X-4.59g/L. DBS-Tg was stable for 15 weeks stored at -20 degrees C. Within-subject variability in DBS-Tg was 21.1%. Reagents and antibodies costs for the new serum and DBS assays are approximate to US$1. Conclusions: These new low-cost serum and DBS-Tg assays perform well over a wide range of Tg concentrations, and the field-friendly DBS assay may be particularly useful in population studies of iodine nutrition.