Sensitive enzyme immunoassay with beta-D-galactosidase-Fab conjugate for detection of type A influenza virus antigen in clinical specimens.

Journal of Clinical Microbiology
M W HarmonS Z Wilson

Abstract

The most sensitive method for diagnosis of type A influenza virus infection is isolation of the agent in cell culture. However, detection and identification may require several days to complete. This delay in diagnosis prevents effective use of the antiviral agents available for treatment of type A influenza infection. As a rapid diagnostic method, enzyme immunoassay (EIA) is attaining increased usage for direct detection of viral antigen in clinical specimens. Standard EIA techniques, however, are usually not sensitive enough for reliable detection of viral antigen in respiratory secretions. We developed a conjugate consisting of the antigen-binding fragment of goat antirabbit immunoglobulin G coupled to beta-d-galactosidase, using the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. Other immunoreagents in our EIA consisted of guinea pig and rabbit antisera to influenza A/Brazil/11/78 (H1N1) for microtiter plate coating and primary antiserum, respectively. The sensitivity of this EIA was tested with 60 clinical specimens containing influenza A/England/333/80 (H1N1) which closely resembles A/Brazil. Of 31 initial specimens, collected within 24 h of the onset of symptoms, 27 (87%) were positive, using a ...Continue Reading

References

Sep 1, 1979·Journal of Clinical Microbiology·R H Yolken, P J Stopa
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Citations

Aug 1, 1988·Journal of Medical Virology·J D CoonrodJ C Donofrio
Jan 1, 1989·Microbiology and Immunology·T KitagawaH Ogura
Jan 1, 1987·Microbiology and Immunology·T KitagawaS Mogi
Jan 1, 1994·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·J C DonofrioT M Chambers
Mar 1, 1984·Journal of Clinical Microbiology·J D CoonrodL C Hsu

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