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Clin Chim Acta. 2015 Jul 20;447:90-5. doi: 10.1016/j.cca.2015.05.010. Epub 2015 May 22.

Bridging-type enzyme-linked immunoassay for zinc transporter 8 autoantibody measurements in adult patients with diabetes mellitus.

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Diabetes Research Group, Institute of Life Sciences, Swansea University, Swansea SA2 8PP, UK. Electronic address:
FIRS Laboratories, RSR Ltd, Cardiff CF14 5DU, UK.
Institute of Molecular and Experimental Medicine, Cardiff University, Cardiff CF14 4YU, UK.
Diabetes Research Group, Institute of Life Sciences, Swansea University, Swansea SA2 8PP, UK.



A bridging-type ELISA for measuring autoantibodies to zinc transporter 8 (ZnT8A) was assessed using samples from different forms of diabetes mellitus.


ZnT8A were measured using an ELISA in patients with type 1 diabetes mellitus (T1DM; n=94), latent autoimmune diabetes of adulthood (LADA; n=51), type 2 diabetes mellitus (T2DM; n=59) and healthy blood donors (HBD; n=200). ZnT8A in ELISA and immunoprecipitation assays (IPA) using ZnT8 dimer (W325/R325) and monomers (W325, R325 and Q325) were compared.


Inter- and intra-assay coefficients of variation (CV) were 7.1% and 1.7%, respectively (medium ZnT8A) and 8.5% and 2.7%, respectively (high ZnT8A). In the ELISA 51/94 (54.3%) T1DM, 16/51 (31.4%) LADA and 1/59 (1.7%) T2DM sera were ZnT8A positive. ROC analysis of T1DM and HBD for the ELISA showed 54% sensitivity and 99% specificity (cutoff 15u/mL) and AUC 0.80 (95% CI, 0.74-0.86). ELISA and IPA measurements were in very good agreement (r=0.856, k=0.889, n=204). Measurement of ZnT8A in addition to autoantibodies for GAD, IA-2 and insulin increased antibody positivity in T1DM by 4.3%, from 80.9% to 85.1%.


The bridging-type ELISA is a convenient and reproducible method for determination of ZnT8A in serum. Measurement of ZnT8A increased autoantibody positivity in adult T1DM.


Autoantibody; ELISA; Type 1 diabetes; ZnT8A

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