Rapid detection of respiratory syncytial virus and influenza A virus in cell cultures by immunoperoxidase staining with monoclonal antibodies.
Peroxidase-labeled monoclonal antibodies against respiratory syncytial virus (RSV) and influenza A virus were used for immunoperoxidase staining (IPS) of cell cultures inoculated with nasopharyngeal aspirates. Cells were grown in 24-well plates, and specimens were inoculated by low-speed centrifugation. Cultures were incubated for 2 days at 37 degrees C and then fixed, stained, and observed by light microscopy. IPS was compared with standard virus isolation by using cultures of human diploid fibroblasts and Vero, HEp-2, and HeLa cell lines for RSV and Madin-Darby canine kidney cells for influenza A virus; these cultures were inoculated with specimens that were previously stored at -70 degrees C. Of 40 known RSV-positive specimens, 30 were found to be positive on reinoculation by both methods, and an additional 5 specimens were found to be positive by IPS only. Of 190 specimens tested for influenza A virus, 14 were positive by IPS and in tubes, and a further 8 specimens were positive by IPS only. IPS was also compared with direct detection of viral antigens in nasopharyngeal aspirates by a time-resolved fluoroimmunoassay (TR-FIA). Fresh nasopharyngeal aspirates were inoculated into human diploid fibroblasts and Madin-Darby canine kidney cells and tested for RSV and influenza A virus, respectively, by IPS. Of 110 specimens tested for RSV, 37 were positive in total, 32 were positive by IPS, and 33 were positive by TR-FIA. Of 150 specimens tested for influenza A virus, 39 were positive in total, 35 were positive by IPS, and 34 were positive by TR-FIA. IPS of cultures inoculated by centrifugation and incubated for 2 days is a sensitive method for the diagnosis of respiratory virus infections, and 24-well plates allow for the easy processing of a large number of specimens.
Department of Virology, University of Turku, Finland.
M
M
Waris
T
T
Ziegler
M
M
Kivivirta
O
O
Ruuskanen
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis*; D004196:Disease Outbreaks; D004285:Dogs; D015200:Fluoroimmunoassay; D005602:France / Q000453:epidemiology; D006367:HeLa Cells; D006801:Humans; D007124:Immunoenzyme Techniques; D009980:Influenza A virus / Q000276:immunology / Q000302:isolation & purification*; D007251:Influenza, Human / Q000150:complications / Q000453:epidemiology* / Q000276:immunology; D009305:Nasopharynx / Q000276:immunology*; D012136:Respiratory Syncytial Viruses / Q000276:immunology / Q000302:isolation & purification*; D010253:Respirovirus Infections / Q000150:complications / Q000453:epidemiology* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral