ADAMTS13 Activity Measurement by ELISA and Fluorescence Resonance Energy Transfer Assay.

Abstract

Accurate estimation of ADAMTS13 (a disintegrin-like and metalloprotease with thrombospondin type 1 motif, member 13) activity level is crucial in the diagnostic setting of differentiation between thrombotic thrombocytopenic purpura (TTP) and other thrombotic microangiopathies. The original assays were too cumbersome and time-consuming for use in the acute situation, and treatment was often based on clinical findings alone, with confirmatory laboratory assays following days or weeks later. Rapid assays are now available that can generate results fast enough to impact on immediate diagnosis and management. Assays based on fluorescence resonance energy transfer (FRET) or chemiluminescence principles can generate results in less than an hour, although they require specific analytical platforms. Enzyme-linked immunosorbent assays (ELISA) can generate results in about 4h, but do not require specialized equipment beyond ELISA plate readers that are in regular use in many laboratories. The present chapter describes principles, performance, and practical aspects of an ELISA and a FRET assay, for quantitative measurement of ADAMTS13 activity in plasma.