Abstract
This study investigated the influence of viral interference on the detection of enteric viruses using the integrated cell culture (ICC)-PCR with a BGM cell line. It was possible to detect 102 plaque-forming units (PFU)/flask of enterovirus 71 (EV71) in spite of the presence of 104 PFU/flask of adenovirus 40 (AdV40). Meanwhile, 104 PFU/flask of AdV40 was not detected in the presence of 102 PFU/flask of EV71. This inhibition of AdV40 detection using ICC-PCR was attributable to the growth of EV71, because the addition of a growth inhibitor of EV71 (rupintrivir) neutralized the detection inhibition of AdV40. The growth inhibition of AdV40 under co-infection with EV71 is probably caused by the immune responses of EV71-infected cells. AdV is frequently used as a fecal contamination indicator of environmental water, but this study demonstrated that false-negative detection of infectious AdV using ICC-PCR could be caused by the co-existence of infectious EV in a water sample. The addition of rupintrivir could prevent false-negative detection of AdV using ICC-PCR. This study, therefore, emphasizes the importance of confirming the presence of multiple enteric viruses in a sample derived from environmental water prior to the application of ICC-PCR because the viral interference phenomenon may lead to the false-negative detection of target viruses.
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Acknowledgements
We thank Dr. Hiroyuki Shimizu and Dr. Minetaro Arita, National Institute of Infectious Diseases, Japan for providing viral strains and suggestions on growth inhibitors used in this study.
Funding
This study was funded by Japan Society for the Promotion of Science KAKENHI 16 K12635 (Grant-in-Aid for Challenging Exploratory Research).
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Conceptualization, D.S., M.K., and S.O.; methodology, D.S. and R.W.; formal analysis, W.O.; investigation, R.W. and W.O.; resources, D.S.; data curation, R.W. and W.O.; writing—original draft preparation, D.S.; writing—review and editing, D.S., M.K., M.A., and S.O.; visualization, D.S.; supervision, D.S.; project administration, D.S.; funding acquisition, D.S.”
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Sano, D., Watanabe, R., Oishi, W. et al. Viral Interference as a Factor of False-Negative in the Infectious Adenovirus Detection Using Integrated Cell Culture-PCR with a BGM Cell Line. Food Environ Virol 13, 84–92 (2021). https://doi.org/10.1007/s12560-020-09453-x
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DOI: https://doi.org/10.1007/s12560-020-09453-x