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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.

Author information

1
Diabetes Research Group, Institute of Life Sciences, Swansea University, Swansea SA2 8PP, UK. Electronic address: G.J.Dunseath@swansea.ac.uk.
2
FIRS Laboratories, RSR Ltd, Cardiff CF14 5DU, UK.
3
Institute of Molecular and Experimental Medicine, Cardiff University, Cardiff CF14 4YU, UK.
4
Diabetes Research Group, Institute of Life Sciences, Swansea University, Swansea SA2 8PP, UK.

Abstract

AIMS:

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

METHODS:

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.

RESULTS:

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%.

CONCLUSIONS:

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.

KEYWORDS:

Autoantibody; ELISA; Type 1 diabetes; ZnT8A

PMID:
26006309
DOI:
10.1016/j.cca.2015.05.010
[Indexed for MEDLINE]

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