Despite the clinical importance of influenza virus in pediatric respiratory infections, the optimal set of diagnostic tests to use when conducting studies using archival samples is not clear. In this study, we compared diagnostic tests for influenza virus in 75 children younger than 5 years of age who presented with symptomatic respiratory infection during one of four influenza seasons, had negative viral cultures for other respiratory pathogens, and had both an archival nasal aspirate obtained at the time of illness and serology spanning that influenza season. For all eligible children, we compared the results of viral culture performed at the time of collection with serology and PCR of archival nasal aspirates. Using real-time viral culture as the "gold standard," the test characteristics of PCR of archival nasal aspirates (sensitivity, 82%; specificity, 100%) and serology (sensitivity, 82%; specificity, 87%) were similar. The relatively low sensitivity of PCR of archival nasal samples in this study compared to that of PCR of fresh samples in a previous study suggests that RNA degradation occurred despite storage of the specimens at ؊70°C. RNA degradation would also explain why only 11 (52%) of 21 archival nasal samples that had positive influenza virus cultures at the time of collection had positive repeat cultures in the summer of 2000. Thus, in archival specimens stored at ؊70°C, PCR was more sensitive than viral culture. However, testing of fresh specimens had the highest yield in this study. Studies of optimal methods for specimen storage are needed.Influenza virus is an important cause of febrile respiratory infections in children. Large population-based studies using administrative databases indicate that rates of hospitalization, antibiotic use, and outpatient visits for children consistently increase when influenza virus is circulating (8,10). Prospective studies of children in inpatient and outpatient settings using viral culture alone or in combination with serology indicate that a high proportion of respiratory illness during the winter is caused by influenza virus (1,3,5,6,11,21). However, estimates of the medical care burden of influenza virus in children have been uncertain, due in part to the lack of large population-based studies that have used state-of-the-art viral diagnostic tests. In fact, the optimal set of diagnostic tests to detect influenza virus infections is debatable, particularly when retrospective epidemiologic studies are conducted using archival samples.The Centers for Disease Control and Prevention (CDC) recently established a new population-based surveillance network, the New Vaccine Surveillance Network (NVSN), to determine the burden of viral respiratory illnesses in hospitalized children younger than 5 years of age. In a previous NVSN study, the results of a rapid diagnostic test for influenza virus were compared to those of viral culture and/or PCR (13). In that study a fully validated and Clinical Laboratory Improvement Amendments-approved PCR of freshly collected samples...
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