Multiplex agglutination-PCR (ADAP) autoantibody assays compared to radiobinding autoantibodies in type 1 diabetes and celiac disease

Childhood screening for type 1 diabetes comparing automated multiplex Antibody Detection by Agglutination-PCR (ADAP) with single plex islet autoantibody radiobinding assays

“…Previously, we have reported the 5-plex ADAP method for detection of four islet autoantibodies and tissue transglutaminase autoantibodies on an automated Hamilton MicroLab STAR systems. 30 Briefly, 4 μL serum or plasma samples were incubated with 8 μL DNA-barcoded autoantigens at 37 °C for 30 min. Heparin plasma was avoided as heparin is a well-known inhibitor of the PCR.…”

“… 24 Prior attempts to replace the RBA with multiplex autoantibody assays include two-sided ELISA for GADA, IA-2A and ZnT8A, 25 , 26 luciferase immunoprecipitation system (LIPS), 27 multiplex assay by electrochemiluminescence (ECL) 28 and the Antibody Detection by Agglutination-PCR (ADAP) assay. 29 , 30 , 31 , 32 …”

“…We have previously compared the automated multiplex ADAP assay with the gold standard RBA in 500 samples from patients newly diagnosed with type 1 diabetes and healthy controls. 30 It was concluded that the simultaneous analysis of 5 different autoantibodies by ADAP in sample volume reduced to only 4 μL and at an increased lower detection limit makes the ADAP automated autoantibody assay advantageous for high throughput screening. 30 The threshold levels are critical in any screening to control the false positive rate.…”

“… 30 It was concluded that the simultaneous analysis of 5 different autoantibodies by ADAP in sample volume reduced to only 4 μL and at an increased lower detection limit makes the ADAP automated autoantibody assay advantageous for high throughput screening. 30 The threshold levels are critical in any screening to control the false positive rate. Our analysis was therefore extended to 2300 patients, 1–10 years of age, with newly diagnosed stage 3 type 1 diabetes 7 , 33 , 34 and 2495 controls to establish diagnostic accuracy by automated multiplex ADAP compared to standard RBA.…”

Effective assay technologies fit for large-scale population screening of type 1 diabetes

“…The limitation of the ECL assay platform is the requirement for a special plate reader. A new multiplex agglutination-PCR (ADAP) autoantibody assay method was reported very recently ( 30 ) and it combines four IAbs and TGA in one single well. The ADAP assay achieved good sensitivity and specificity in a study with a group of T1D patients vs healthy controls, however improvement on IA-2A and ZnT8A is still needed according to the data reported.…”