TY - JOUR
T1 - Bridging-type enzyme-linked immunoassay for zinc transporter 8 autoantibody measurements in adult patients with diabetes mellitus
AU - Dunseath, Gareth
AU - Ananieva-Jordanova, Rossitza
AU - Coles, Rebecca
AU - Powell, Michael
AU - Amoroso, Marie
AU - Furmaniak, Jadwiga
AU - Smith, Bernard Rees
AU - Dayan, Colin
AU - Luzio, Stephen
N1 - Publisher Copyright:
© 2015 Elsevier B.V.
PY - 2015/7/1
Y1 - 2015/7/1
N2 - 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 15. u/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.
AB - 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 15. u/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.
KW - Autoantibody
KW - ELISA
KW - Type 1 diabetes
KW - ZnT8A
UR - https://www.scopus.com/pages/publications/84931281647
U2 - 10.1016/j.cca.2015.05.010
DO - 10.1016/j.cca.2015.05.010
M3 - Article
C2 - 26006309
AN - SCOPUS:84931281647
SN - 0009-8981
VL - 447
SP - 90
EP - 95
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
ER -
