Development of Techniques for Evaluating the Virus Removal Rate using Adenovirus

아데노바이러스를 이용한 바이러스 제거율 평가를 위한 기법 개발

  • Received : 2015.09.24
  • Accepted : 2015.10.30
  • Published : 2015.12.15


Waterborne infectious disease is induced by several pathogenic microbes such as bacteria, viruses and protozoans, and the cases caused by viral infection is currently increasing. Water treatment process could reduce the number of virus in the water, but there were many difficulties to completely remove the virus particles from water. Therefore, the membrane separation technology which was reported to effectively remove pollutants from raw water has attracted increasing attention and demand. Since its efficiency has been introduced, demands for evaluation method toward the membrane filtration process are increasing. However, progression of the method development is slow due to the difficulties in cultivation of several waterborne viruses from animal models or cell culture system. To overcome the difficulties, we used adenovirus, one of the commonly isolated pathogenic waterborne viruses which can grow in cell culture system in vitro. The adenovirus used in this study was identified as human adenovirus C strain. The adenovirus was spiked in the raw water and passed through the microfiltration membrane produced by Econity, a Korean membrane company, and then the viral removal rate was evaluated by real-time PCR. In the results, the amount of virus in the filtered water was decreased approximately by 5 log scale. Because coagulant treatment has been known to reduce filtering function of the membrane by inducing fouling, we also investigated whether there was any interference of coagulant. In the results, we confirmed that coagulant treatment did not show significant interference on microfiltration membrane. In this study, we found that waterborne virus can be effectively removed by membrane filtration system. In particular, here we also suggest that real-time PCR method can rapidly, sensitively and quantitatively evaluate the removal rate of virus. These results may provide a standard method to qualifying membrane filtration processes.


Adenovirus;Microfilteration membrane;Real-time PCR;Waterborne viruses


  1. Waterworks Law,, lsInfoP&lsId=001818#0000 (June 4, 2015).
  2. Abbaszadegan, M., Stewart, P., LeChevallier, M. (1999). A strategy for detection of viruses in groundwater by PCR, Appl Environ Micorbiol., 65, 444-449.
  3. Ahn, H.Y. (2007). Case studies and R&D on membranes in K-water, J Kor Soc Environ Eng., 29, 1318-1325.
  4. Allard, A., Girones, R., Juto, P., Wadell, G. (1990). Polymerase chain reaction for detection of Adenoviruses in stool samples, J Clin Microbiol., 28, 2659-2667.
  5. Beller, M., Ellis, A., Lee, S.H., Drebot, M.A., Jenkerson, S.A., Funk, E., Sobsey, M.D., Simmons III, O.D., Monroe, S.S., Ando, T., Noel, J., Petric, M., Middaugh, J.P. and Spika, J.S. (1997). Outbreak of viral gastroenteritis due to a contaminated well. International consequences, JAMA., 278, 563-568.
  6. Bitton, G., Farrah, S.R., Montague, C.L., Akin, E.W. (1986). Viruses in drinking water, Environ Sci Technol., 20, 216-222.
  7. Cheryan, M. (1998). Ultrafiltration and microfiltration handbook 2nd Ed., Technoimic Publishing Company, New York.
  8. Cho, H.B., Lee, S.H., Cho, J.C., Kim, S.J. (2000). Detection of adenoviruses and enteroviruses in tap water and river water by reverse transcription multiplex PCR, Can J Microbiol., 46, 417-424.
  9. Cukor, G., Blacklow, N.R. (1984). Human viral gastroenteritis, Microbiol Rev., 48, 157-179.
  10. Defrance, L., Jaffrin, M.Y. (1999). Reversibility of fouling formed in activated sludge filtration, J Membrane Sci., 157, 73-84.
  11. Dubois, E., Merle, G., Roquier, C., Trompette, A.L., Le Guyader, F., Cruciere, C., Chomel, J.J. (2004). Diversity of enterovirus sequences detected in oysters by RT-heminested PCR, Int J Food Microbiol., 92, 35-43.
  12. Duizer, E., Bijkerk, P., Rockx, B., De Groot, A., Twisk, F., Koopmans, M. (2004). Inactivation of caliciviruses, Appl Environ Microbiol., 70, 4538-4543.
  13. Fong, T.T., Lipp, E.K. (2005). Enteric viruses of humans and animals in aquatic environments: health risks, detection, and potential water quality assessment tools. Microbiol Mol Biol Rev., 69, 357-371.
  14. Fout, G.S., Martinson, B.C., Moyer, M.W., Dahling, D.R. (2003). A multiplex reverse transcription-PCR method for detection of human enteric viruses in groundwater. Appl Environ Microbiol., 69, 3158-3164.
  15. Gilgen, M., Wegmuller, B., Burkhalter, P., Buhler, H.P., Muller, U., Luthy, J., Candrian, U. (1995). Reverse transcription PCR to detect enteroviruses in surface water. Appl Environ Microbiol., 61, 1226-1231.
  16. Gu, Z., Belzer, S.W., Gibson, C.S., Bankowski, M.J., Hayden, R.T. (2003). Multiplexed, real-time PCR for quantitative detection of human adenovirus. J Clin Microbiol ., 41, 4636-4641.
  17. Harris, J.R., Cohen, M.L., Lippy, E.C. (1983). Water-related disease outbreaks in the United States, 1981. J Infect Dis., 148, 759-762.
  18. Hur, H.C., Rhee, O.J., Lee, K.J., Kim, K.H., Choi, Y.J., Lee, J.H., Hong, S.W. (2008). Analysis of Membrane Integrity and Removal Efficiency Considering Membrane Defect and Pore size. J Korean Soc Water Qual., 24, 423-429.
  19. Jung, E.Y., Park, H.G., Cha, D.J., Jung, M.E., You, P.J. (2009). Removal Efficiency in Water Treatment Process and Characteristic of Cell Sensitivity of Waterborne Enteric Viruses, J Life Sci., 19, 373-377.
  20. Knolle, H. (1995). Transmission of poliomyelitis by drinking water and the problem of prevention, Gesundheitswesen, 57, 351-354.
  21. Madaeni, S.S., Fane, A.G., Grohmann, G.S. (1995). Virus removal from water and wastewater using membrane, J Membrane Sci., 102, 65-75.
  22. Maunula, L., Miettinen, I.T., Von Bonsdorff, C.H. (2005). Norovirus outbreaks from drinking water, Emerg Infect Dis., 11, 1716-1721.
  23. Papaventsis, D., Siafakas, N., Markoulatos, P., Papageorgiou, G.T., Kourtis, C., Chatzichristou, E., Economou, C., Levidiotou, S. (2005). Membrane adsorption with direct cell culture combined with reverse transcription-PCR as a fast method for identifying enteroviruses from sewage, Appl Environ Microbiol., 71, 72-79.
  24. Schwab, K.J., De Leon, R., Sobsey, M.D. (1993). Development of PCR methods for enteric virus detection in water, Wat Sci Tech., 27, 211-217.
  25. Taedosiu, C.C., Kennedy, M.D., Straten, H.A., Schippers, J.C. (1999). Evaluation of secondary refinery effluent treatment using ultrafiltration membranes, Water Resarch., 33, 2172-2180.
  26. Vainio, K., Myrmel, M. (2006). Molecular epidemiology of norovirus outbreaks in Norway during 2000 to 2005 and comparison of four norovirus real-time reverse transcriptase PCR assays, J Clin Micorbiol., 44, 3695-3702.


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