RESUMEN
An all-monoclonal antibody, time-resolved fluoroimmunoassay was compared with several enzyme immunoassays for the detection of respiratory syncytial virus and parainfluenza virus type 1, 2, and 3 antigens in clinical specimens. The most sensitive enzyme immunoassay for parainfluenza virus type 1 was an all-monoclonal antibody assay with biotin-labeled detector antibody and streptavidin-peroxidase conjugate, but for respiratory syncytial virus and parainfluenza virus types 2 and 3 the most sensitive assay was a polyclonal antibody assay with horse capture antibodies and bovine or rabbit detector antibodies with anti-species peroxidase. All tests were evaluated with nasopharyngeal aspirate specimens from respiratory illnesses and with cell culture harvests of multiple strains of each virus isolated over many years. The time-resolved fluoroimmunoassay detected respiratory syncytial virus antigen in 92% of the specimens positive by culture, which was a decidedly higher sensitivity than either the monoclonal or polyclonal antibody enzyme immunoassay format (62 and 76%, respectively). For the parainfluenza viruses the time-resolved fluoroimmunoassay detected type-specific antigen in 94 to 100% of culture-positive specimens and again was more sensitive than the all-monoclonal antibody enzyme immunoassays (75 to 89%) or all-polyclonal antibody enzyme immunoassays (66 to 95%). Combined with results from a previously reported adenovirus time-resolved fluoroimmunoassay, these tests identified respiratory antigens in large numbers of clinical specimens.
Asunto(s)
Antígenos Virales/análisis , Virus Sincitiales Respiratorios/aislamiento & purificación , Respirovirus/aislamiento & purificación , Anticuerpos Monoclonales/inmunología , Biotina , Cromatografía por Intercambio Iónico , Técnica del Anticuerpo Fluorescente , Humanos , Técnicas para Inmunoenzimas , Valor Predictivo de las Pruebas , Virus Sincitiales Respiratorios/inmunología , Respirovirus/inmunologíaRESUMEN
A sensitive time-resolved fluoroimmunoassay (TR-FIA), adapted from TR-FIA procedures already described, was developed with monoclonal antibodies and compared with several enzyme immunoassays (EIAs) for detecting adenovirus antigens in clinical specimens. The most sensitive EIA was an all-monoclonal assay with biotin-labeled detector antibody and streptavidin-peroxidase conjugate. All tests were evaluated with nasopharyngeal aspirate specimens from respiratory illness, with tissue homogenates from patients with systemic infection, and with stool specimens from gastrointestinal illnesses. For respiratory and tissue specimens, the TR-FIA detected adenovirus in 85% of the specimens positive by culture, which was a sensitivity similar to those of the all-monoclonal biotin-avidin EIA (79%) and the polyclonal-capture biotin-avidin EIA (88%). For stool specimens, the TR-FIA detected adenovirus in 100% of the specimens positive by culture, which was a decidedly higher sensitivity than either EIA format (78 and 75%, respectively). The TR-FIA was shown to be an efficient, flexible, and specific test for large numbers of clinical specimens.
Asunto(s)
Infecciones por Adenoviridae/diagnóstico , Infecciones por Adenovirus Humanos/diagnóstico , Adenovirus Humanos/inmunología , Antígenos Virales/análisis , Técnica del Anticuerpo Fluorescente , Adenovirus Humanos/aislamiento & purificación , Anticuerpos Monoclonales/inmunología , Anticuerpos Antivirales/inmunología , Niño , Preescolar , Heces/microbiología , Humanos , Técnicas para Inmunoenzimas , Lactante , Nasofaringe/microbiología , Valor Predictivo de las PruebasRESUMEN
Monoclonal antibodies that are broadly reactive with either influenza A or influenza B viruses were used to develop a 2- to 3-h antigen capture time-resolved fluoroimmunoassay (TR FIA) for detecting influenza viral antigens in both original nasopharyngeal aspirate specimens and in tissue cultures inoculated with nose or throat swab specimens. The lower limit of sensitivity of the assay was about 10 pg of protein as determined with purified influenza A nucleoprotein expressed by recombinant DNA. When the TR FIA was performed with 96 nasopharyngeal aspirate specimens collected during outbreaks of influenza A (H3N2) virus and the results were compared with serodiagnosis results with paired sera, the specificity and sensitivity of TR FIA for the demonstration of influenza A infections were 95 and 85%, respectively. In culture confirmation assays, more than 80% of the swab specimens that grew influenza A or B virus within 7 days could be identified by the TR FIA within 48 h of the inoculation of cells. The results are consistent with those previously reported for respiratory syncytial virus and extend the applicability of monoclonal antibody-based TR FIA for the rapid diagnosis of acute respiratory viral infections.