New Sandwich-Type Enzyme-Linked Immunosorbent Assay for Human MxA Protein in a Whole Blood Using Monoclonal Antibodies Against GTP-Binding Domain for Recognition of Viral Infection

Objectives To develop a clinically significant and practical enzyme‐linked immunosorbent assay (ELISA) for the detection of MxA protein in human whole blood, a biological marker of viral infection. Design and Methods A sandwich ELISA suitable for the measurement of human MxA protein in whole blood was developed using mouse monoclonal antibodies (mAbs) raised against the GTP‐binding domain of human MxA protein. Prior to the assay, the whole blood sample was treated with special buffer to extract the MxA protein, improve its stability, and avoid interference from hemoglobin. Results This ELISA meets all the requirements for use in routine clinical assays, especially in terms of sensitivity (detection limit: 1.3 ng/ml whole blood), accuracy (recovery: 93.0–100.0%), and rapidity (