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A biotin-enhanced enzyme immunoassay (EIA) for respiratory syncytial virus (RSV) antigen detection (TESTPACK RSV®) was prospectively compared with virus isolation in cell culture and immunofluorescence. Of 156 nasopharyngeal swab specimens from infants with respiratory symptoms, 81 (52%) yielded RSV in culture. Compared with culture the sensitivity of the EIA was 95% and specificity was 92%; the specificity increased to 97% with a blocking assay. Compared with immunofluorescence the sensitivity of EIA was 92% and specificity was 93%. In order to assess the performance of TESTPACK RSV® as a bedside test, nasopharyngeal swabs from 49 children were tested by EIA at the bedside by housestaff and by immunofluorescence in the laboratory; the sensitivity of the EIA was lower (78%) while specificity remained high (95%). Inclusion of older children may have resulted in diminished sensitivity. The TESTPACK RSV® is a simple, rapid test that performs well and is easily adaptable to an office setting. Further evaluation of the test in older children may be required.