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Detection of Dual-Infected Cases of Adenoviruses and Coxsackieviruses Type B by Real-Time PCR but not by the Conventional Viral Culture Technique by Tsuguto Fujimoto, Michiyo Shinohara, Masahiro Ito, Teruo Okafuji, Takao Okafuji, Osamu Nishio, Hiromu Yoshida, Hideaki Shimizu, Masatsugu Chikahira, Gia Tung Phan, Hiroshi Ushijima

The aim of this study was to evaluate the applicability of diagnostic methods for dual-infected cases of human adenoviruses (AdVs) and coxsackieviruses type B (CBs). For this purpose, 100 nasopharyngeal samples from patients with acute exudative tonsillitis and clinically suspected AdV infection were analyzed. Using PCR and real-time PCR techniques for AdVs and CBs, we found 86 AdVs-only positive samples; we also found five dualinfected samples containing 5.4×105 to 7.1×108 copies/mL of AdV genomes and 1.4×104 to 1.3×109 copies/mL of CB genomes. By viral culture using A549 cells, two co-infected samples, which contained over 108 copies/mL of AdV genomes and <105 copies/mL of CB genomes, became AdV dominant, while three samples with less than 2.0×106 copies/mL of AdV genomes became CB dominant. An immunochromatography kit for diagnosing AdVs at the bedside was positive for 3/5 dual-infected patients, and PCR techniques for AdVs and CBs were both positive for 5/5. Viral culture is usually considered to be the gold standard for AdV diagnosis, but our results demonstrate the importance of PCR applications for the detection of AdV and CB genomes, particularly in clinical cases of suspected AdV infection. Even though the sample size of dual infection (n=5) is small, our results show the existence of dual infection cases which were difficult to diagnose by viral culture alone.

DOI: Clin. Lab. 2007;53:605-609