J Immunol Methods. 2016 Jan 22. pii: S0022-1759(16)30010-2. doi: 10.1016/j.jim.2016.01.011. [Epub ahead of print]
A multiplex assay combining insulin, GAD, IA-2 and transglutaminase autoantibodies to facilitate screening for pre-type 1 diabetes and celiac disease.
Abstract
At the current time, multiple candidate interventions are being proposed to abrogate or slow progression to type 1 diabetes
(T1D) among islet autoantibody (iAb) positive subjects, but mass
screening for eligible subjects and the general population remains a
laborious and inefficient process. We have recently developed and
extensively validated nonradioactive iAb assays using
electrochemiluminescense (ECL) detection with an excellent sensitivity
and specificity compared to the gold-standard radioassays. Using ECL
detection on a platform from MesoScale Discovery (MSD) allows the
measurement of four antibodies
in a single well using a small blood volume (6ul). In the present study
using a MSD QuickPlex 4-Spot plate, we successfully combined three iAb
to insulin (IAA), GAD65 (GADA), and IA-2 (IA-2A) with tissue transglutaminase
autoantibodies (TGA) in a single well of a 96 well plate. We tested 40
new onset T1D patients, all positive for at least one iAb and a half of
them positive for TGA by radioassay, as well as 50 healthy controls. The
multiplex assay retained 100% sensitivity and 100% specificity for all
four autoantibodies in terms of positivity identified in patients versus
normal controls compared to the corresponding standard radioassays and
our single ECL assays. The multiplex ECL assay was able to identify more
positivity than current radioassays for IAA and TGA. The development of
this multiplex assay will facilitate high-throughput screening for T1D
and celiac disease risk in the general population.
Copyright © 2015. Published by Elsevier B.V.
Copyright © 2015. Published by Elsevier B.V.
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