Microfluidic Electrochemical Biosensor for the Detection of Platelet Factor 4 Antibody‐Mediated Thrombotic Disorders

Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a new disorder that emerged in the wake of coronavirus disease 2019 (COVID-19) vaccination. It is a rare but life-threatening condition that requires an aggressive course of treatment to improve patient outcomes. Currently, the diagnosis of VITT relies on a comprehensive panel of criteria, including a history of recent vaccination, platelet count, positive enzyme linked immunosorbent assays (ELISA) result for a closely related thrombotic disorder, heparin-induced thrombocytopenia (HIT), and platelet factor 4 (PF4)-dependent functional assays. The study describes a technically simple antigenic assay designed to detect autoimmune antibodies associated with VITT. It is first shown that cross-linked platelet factor 4 (c-PF4) represents an antigenic target specific for VITT/VITT-like Abs. This antigenic target is then incorporated into a microfluidic electrochemical biosensor and specific and sensitive detection of VITT/VITT-like antibodies is demonstrated in a fully automated manner while using microliter volumes of patient sera. Fifty-five serum samples are tested using the microfluidic electrochemical biosensor and demonstrated high accuracy for the detection of VITT sera and VITT-like monoclonal gammopathy of thrombotic ignificance (MGTS) antibodies compared to healthy controls and HIT patients. It is envisioned that the microfluidic electrochemical biosensor will be used in combination with other clinical criteria to enable timely, sensitive, and specific diagnosis of VITT.