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A novel radioligand binding assay to determine diagnostic accuracy of isoform-specific glutamic acid decarboxylase antibodies in childhood IDDM

Grubin, C. E. ; Daniels, T. ; Toivola, B. ; Landin-Olsson, M. LU ; Hagopian, W. A. ; Li, L. ; Karlsen, A. E. ; Boel, E. ; Michelsen, B. and Lernmark, Å LU (1994) In Diabetologia 37(4). p.344-350
Abstract

Insulin-dependent diabetes mellitus (IDDM) is associated with autoreactivity against GAD but the diagnostic sensitivity (positivity in disease) and specificity (negativity in health) of isoform-specific GAD antibodies have yet to be defined in assay systems suitable for screening large number of samples. One set of IDDM patient (n=10) and control (n=50) standard sera were used to develop quantitative antibody assays with in vitro synthesized recombinant 35S-methionine-labelled GAD65 and GAD67, respectively, and protein A-Sepharose to separate free from antibody-bound ligand. Binding levels were not normally distributed (p<0.0001) and therefore, the diagnostic accuracy of GAD antibodies was analysed by the ROC plots in... (More)

Insulin-dependent diabetes mellitus (IDDM) is associated with autoreactivity against GAD but the diagnostic sensitivity (positivity in disease) and specificity (negativity in health) of isoform-specific GAD antibodies have yet to be defined in assay systems suitable for screening large number of samples. One set of IDDM patient (n=10) and control (n=50) standard sera were used to develop quantitative antibody assays with in vitro synthesized recombinant 35S-methionine-labelled GAD65 and GAD67, respectively, and protein A-Sepharose to separate free from antibody-bound ligand. Binding levels were not normally distributed (p<0.0001) and therefore, the diagnostic accuracy of GAD antibodies was analysed by the ROC plots in population-based, consecutively-diagnosed, recent onset, 0-14 year-old patients (n=105), and matched, healthy control subjects (n=157). The ROC plots showed that the diagnostic sensitivity of GAD65 antibodies was 77% and the specificity 92% compared with 8% and 98%, respectively for GAD67 antibodies. In the IDDM sera, GAD65 and GAD67 antibodies were concordant in 7% (6 of 81) and GAD65 antibodies and ICA in 89% (72 of 81) without a correlation between the autoantibody levels. Autoantibodies to recombinant human islet GAD65 are specific and sensitive markers for childhood IDDM in this immunoassay with in vitro synthesized 35S-methioninelabelled recombinant GAD.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
autoimmunity, diabetes mellitus, diagnostic accuracy, Glutamic acid decarboxylase, islet cell antibodies, receiver-operating characteristic plot
in
Diabetologia
volume
37
issue
4
pages
344 - 350
publisher
Springer
external identifiers
  • scopus:0028258006
  • pmid:8063033
ISSN
0012-186X
DOI
10.1007/BF00408469
language
English
LU publication?
yes
id
08363607-a6e7-46d2-9004-8f165f58070e
date added to LUP
2019-09-11 09:06:17
date last changed
2021-03-07 05:28:25
@article{08363607-a6e7-46d2-9004-8f165f58070e,
  abstract     = {<p>Insulin-dependent diabetes mellitus (IDDM) is associated with autoreactivity against GAD but the diagnostic sensitivity (positivity in disease) and specificity (negativity in health) of isoform-specific GAD antibodies have yet to be defined in assay systems suitable for screening large number of samples. One set of IDDM patient (n=10) and control (n=50) standard sera were used to develop quantitative antibody assays with in vitro synthesized recombinant <sup>35</sup>S-methionine-labelled GAD65 and GAD67, respectively, and protein A-Sepharose to separate free from antibody-bound ligand. Binding levels were not normally distributed (p&lt;0.0001) and therefore, the diagnostic accuracy of GAD antibodies was analysed by the ROC plots in population-based, consecutively-diagnosed, recent onset, 0-14 year-old patients (n=105), and matched, healthy control subjects (n=157). The ROC plots showed that the diagnostic sensitivity of GAD65 antibodies was 77% and the specificity 92% compared with 8% and 98%, respectively for GAD67 antibodies. In the IDDM sera, GAD65 and GAD67 antibodies were concordant in 7% (6 of 81) and GAD65 antibodies and ICA in 89% (72 of 81) without a correlation between the autoantibody levels. Autoantibodies to recombinant human islet GAD65 are specific and sensitive markers for childhood IDDM in this immunoassay with in vitro synthesized <sup>35</sup>S-methioninelabelled recombinant GAD.</p>},
  author       = {Grubin, C. E. and Daniels, T. and Toivola, B. and Landin-Olsson, M. and Hagopian, W. A. and Li, L. and Karlsen, A. E. and Boel, E. and Michelsen, B. and Lernmark, Å},
  issn         = {0012-186X},
  language     = {eng},
  month        = {04},
  number       = {4},
  pages        = {344--350},
  publisher    = {Springer},
  series       = {Diabetologia},
  title        = {A novel radioligand binding assay to determine diagnostic accuracy of isoform-specific glutamic acid decarboxylase antibodies in childhood IDDM},
  url          = {http://dx.doi.org/10.1007/BF00408469},
  doi          = {10.1007/BF00408469},
  volume       = {37},
  year         = {1994},
}