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.
Tkácová
M
M
Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovak Republic.
Varecková
E
E
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis*; D005069:Evaluation Studies as Topic; D006801:Humans; D007124:Immunoenzyme Techniques; D009980:Influenza A virus / Q000276:immunology / Q000302:isolation & purification*; D007251:Influenza, Human / Q000473:pathology / Q000821:virology*; D051379:Mice; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000032:analysis*; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins / Q000032:analysis*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins