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Detection of respiratory syncytial virus in nasopharyngeal secretions by 24-well plate precentrifugation assay using a monoclonal antibody against F protein
Authors:Savón C  Goyenechea A  Valdivia A  Chacón D  Cancio R  Angel-Pérez L  González G  Gavilondo J
Institution:Departamento de Virología, Instituto de Medicina Tropical Pedro Kouri, Havana, Cuba. clara@ipk.sld.cu
Abstract:BACKGROUND: Respiratory syncytial virus (RSV) is responsible for 50% of all bronchiolitis and 25% of pneumonia cases during the first month of life. Detection of the RSV antigen by immunofluorescence in exfoliated nasal epithelium or by other methods in nasopharyngeal swabs is useful in the potentially infected patient because results are available within a few hours. In contrast, RSV antigen detection in cell culture may require as much as 3 weeks. METHODS: Three methods for detection of respiratory syncytial virus in 131 clinical respiratory specimens from patients with acute respiratory disease and bronchiolitis were compared utilizing the following: a precentrifugation immunofluorescence assay using Hep-2 cells, indirect immunofluorescence assay, and conventional tube cell culture using Hep-2 cells. RESULTS: Respiratory syncytial virus was identified in 36 specimens by the three methods previously described. The virus was recovered in 41 (31.3%) samples by precentrifugation immunofluorescence assay, 40 (30.5%) were identified by the immunofluorescence technique, and 38 (29.0%) cases were positive by conventional cell culture. The sensitivity of the precentrifugation assay in relation to the immunofluorescence technique was 90%, the specificity 94.5%, and the agreement, 96.2%. A positive predictive value of 90.2% was obtained. Sensitivity, specificity, agreement, and positive predictive values obtained by the precentrifugation assay variant compared to the conventional cell were 90.8%, 94.5%, 93.1%, and 87.8%, respectively. CONCLUSIONS: The precentrifugation immunofluorescence assay method was as sensitive as the remainder of the methods used in our study and represents a valid alternative for rapid detection of respiratory syncytial virus in clinical samples.
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