Comparison of radioimmunoprecipitation with luciferase immunoprecipitation for autoantibodies to GAD65 and IA-2beta.
OBJECTIVE
To compare the sensitivity and specificity of luciferase immunoprecipitation (LIPS) with radioimmunoprecipitation (RIP) for the measurement of autoantibodies to the type 1 diabetes autoantigens glutamic acid decarboxylase 65 (GAD65) and insulinoma-associated protein (IA)-2beta.


METHODS
Sera from 49 type 1 diabetic patients and 100 nondiabetic control subjects from Diabetes Antibody Standardization Program 2007 were used to screen for autoantibodies to GAD65. An additional 200 type 1 diabetic patients and 200 nondiabetic control subjects were used to validate the GAD65 results and screen for autoantibodies to IA-2beta.


RESULTS
LIPS showed equal sensitivity and specificity to RIP for detecting autoantibodies to GAD65 and IA-2beta. Receiver-operating characteristic analysis revealed that the detection of autoantibodies to GAD65 and IA-2beta by LIPS and RIP were not statistically different.


CONCLUSIONS
The LIPS assay does not require the use of radioisotopes or in vitro transcription/translation and is a practical alternative at the clinical level for the RIP assay.
Burbelo
Peter D
PD
Neurobiology and Pain Therapeutics Section, Laboratory of Sensory Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
Hirai
Hiroki
H
Issa
Alexandra T
AT
Kingman
Albert
A
Lernmark
Ake
A
Ivarsson
Sten-A
SA
Notkins
Abner L
AL
Iadarola
Michael J
MJ
D001323:Autoantibodies / Q000032:analysis* / Q000276:immunology; D003922:Diabetes Mellitus, Type 1 / Q000175:diagnosis / Q000276:immunology; D005968:Glutamate Decarboxylase / Q000276:immunology*; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D008156:Luciferases / Q000378:metabolism*; D015531:Radioimmunoprecipitation Assay / Q000379:methods*; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8 / Q000276:immunology*; D012680:Sensitivity and Specificity
D001323:Autoantibodies; D008156:Luciferases; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2