This was the situation inside our study also, where correlation of patients’ symptoms and findings with infection because of different agents revealed just a few associations, mostly in relation to with -capture ELISA may be useful in the selection of antibiotic treatment, because these antibodies are detected in a high proportion of patients during the acute phase of illness. In conclusion, the use of multiple assays, including PCR, may increase sensitivity in documenting infection with different pathogens in school-age children hospitalized with CAP. morbidity among children in developed countries and has a considerable effect on the health care system. In the developing world, the incidence of pneumonia is higher, and this infection is one of the primary causes of death among young children [1]. Detailed information on the etiology of CAP is required for the formulation of treatment recommendations and the introduction of preventive measures. Evaluation of mixed infections and the Dinoprost tromethamine relative importance of each potential pathogen may also contribute to improved understanding of the etiopathogenesis of this infection. However, identifying the cause of a lower respiratory tract infection remains a challenge for a number of reasons: adequate samples are difficult to obtain, and the differentiation between infection and colonization cannot always be made [1, 2]. Furthermore, the different responsible agents require multiple and complicated Dinoprost tromethamine methodologies for their detection. As more diagnostic tests or combinations of them are performed, the number of potential causes directly or indirectly associated with pneumonia increases [1, 2]. has been identified as the most important cause of bacterial pneumonia in children [1, 2]. and are most common causes in school-age children [1, 2], although recent studies have suggested their importance in younger age groups [3C5]. Viruses, Dinoprost tromethamine with the predominance of respiratory syncytial virus (RSV), have been most commonly associated with pneumonia in infants and young children. However, the role of viruses in many respiratory tract diseases has been readdressed in recent studies that use new sensitive methodologies. This is mostly true for human rhinovirus (RV), which for many years has been considered as a uniquely upper-airway pathogen. Nevertheless, the development of sensitive diagnostic techniques has helped identify RV as significantly associated with lower respiratory tract diseases, such as asthma [6], bronchiolitis [7], and CAP [8]. We conducted a 12-month prospective study involving school-age children hospitalized with CAP to thoroughly investigate the CD244 role of viruses, and atypical and common bacteria by means of several detection techniques, including PCR and serological testing. Patients and Methods A total of 75 patients (37 of whom were boys) aged 5C14 years (median age, 86.5 months) who were consecutively admitted to our department with the diagnosis of CAP during 1 calendar year were enrolled in the study. Patients were included if they had fever (temperature, ?37.5C) and an infiltrate visible on a chest radiograph. Children with any chronic underlying disorders were excluded from the study. A standard questionnaire, including demographic data, clinical symptoms and signs, laboratory and radiological findings, treatment, complications, and duration of hospitalization, was filled out for each patient. A serum sample and a nasopharyngeal wash were obtained from each patient within 48 h of admission. The samples were kept frozen at -70C until further study. Forty-five patients returned for follow-up 1 month after discharge, at which time a convalescent-phase serum sample was obtained. The presence of genomic material of the viruses and atypical bacteria in nasopharyngeal wash samples was examined by RNA or DNA extraction, followed by RT-PCR and direct PCR, respectively. PCR reactions with primer sets and conditions specific for RSV A and B, influenza viruses A (H1N1 and H3N2 subtypes) and B, parainfluenza viruses 1, 2, and 3, adenoviruses, human metapneumovirus, and and were performed as previously described [7, 9C11]. A single round and a seminested PCR were used for detection of RV [12, 13]. Primers were synthesized from MWG AG Biotech; Taq DNA polymerase and PCR buffers and supplements were purchased from Bioron. PCR products were electrophoresed on 2% agarose gels, and amplicons were visualized by ethidium bromide staining. When a PCR of nested or seminested type was used, it was conducted in a separate room to avoid intrasample contamination. cDNAs obtained from viral lysates served as positive controls, whereas several negative controls (virus transport medium or water) were included in each run to exclude the possibility of contamination. IgG, IgA, and IgM antibodies against were measured by an in-house microimmunofluorescence method by means of purified, formalized elementary bodies, with strain K6 as antigen, as described elsewhere [14]. Diagnosis was based on a 4-fold increase in titer between paired serum samples or on the presence of IgM in any serum sample. A commercial immunoassay (Labsystems Oy) was also used for the detection of IgG Dinoprost tromethamine and IgM antibodies to (pneumolysin and C-polysaccharide), were.