A sensitive one-step immunocapture EIA for rapid diagnosis of influenza A

A highly sensitive one-step immunocapture EIA for the detection of influenza A virus antigen directly in a clinical specimen was developed. The sensitivity was achieved by using two high-affinity cross-reactive influenza type A-specific monoclonal antibodies, recognizing independent nonoverlapping epitopes on the influenza A nucleoprotein. One of the two MAbs was used as a capture antibody, while the other was coupled with enzyme peroxidase and served as a detector. Sensitivity to detection of highly purified recombinant influenza A virus nucleoprotein by EIA reached approximately 10 pg. Fifteen purified human influenza A virus strains of H1, H2 and H3 subtypes, isolated during the period 1934-1992, were tested by this system. All the influenza A viruses tested positive, whereas two influenza B viruses used as a control were negative. The efficiency of the system for detection of influenza A viral antigen directly in clinical specimens was confirmed by testing nasal and nasopharyngeal washes and aspirates, tested previously by time-resolved fluoroimmunoassay and by virus culture confirmation assay.