Journal of fish pathology (한국어병학회지)

The Korean Society of Fish Pathology (한국어병학회)
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Genetic analysis of norovirueses in Busan

부산지역 노로바이러스의 유전적 분석

  • Kim, Kwang-Il (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Jin, Ji-Woong (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Jeong, Hyun-Do (Department of Aquatic Life Medicine, Pukyong National University)
  • 김광일 (부경대학교 수산생명의학과) ;
  • 진지웅 (부경대학교 수산생명의학과) ;
  • 정현도 (부경대학교 수산생명의학과)
  • Received : 2011.09.27
  • Accepted : 2011.11.30
  • Published : 2011.12.31
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Abstract

For detection of noroviruses (NVs), we compared various PCR primer sets based on reverse transcription nested PCR (RT-nested PCR) in the water samples from Dong brook in Busan, South Korea. We designed various new primer sets based on the most conserved sequences of the capsid protein gene that react with diverse NVs found in Korea. Designed primer sets (KG1F/KG1R and KG2F/KG2R, named as PNK) for the respective genogroups of NVs, genogroup I and II (GI and GII), were applied to detect NVs in the water samples from Dong brook concentrated with ultracentrifugation. In the application to the water samples, proportion of GI (76.47%) and GII (70.59%) in water samples of Dong brook in RT-nested PCR with the primer sets of this study. However, no significant differences of the proportion of the positive samples were not found between RT-nested PCRs with reported and newly designed primer sets. From the nucleotide sequencing, GI and GII of NVs present in Dong brook were appeared to be the members of 1/2/4/5/9/10 genotypes, and 3/4/5/11/13 genotypes respectively. Appeared genotype 4 of GII known as an one of main genotype found in patients of many Asian countries warned us to consider the risks of norovirus in aquatic environments in southern part of Korea.

본 연구에서는 RT-nested PCR을 수행하여 부산 도심의 하천 중 동천에 존재하는 노로바이러스를 검출하고자 하였다. 기존에 보고되어진 노로바이러스의 capsid protein의 염기서열을 비교하여 노로바이러스 genogroup I,II (GI,II) 를 검출하기 위한 새로운 degenerated primer sets (PNK, KGIF/KGIR and KG2F/KG2R) 를 제작하였으며, 채수한 동천 시료를 초고속원심분리기를 통해 농축 후 물 속에 존재하는 노로바이러스의 검출을 시도하였다. 노로바이러스를 검출하기 위해 PCR primer를 비교한 결과 본 연구에서 제작한 capsid protein gene을 target으로 하는 primer set가 기존에 보고되어 있는 primer set보다 동일 시료에 대한 검출빈도가 우수하였다. 동천에 존재하는 노로바이러스의 오염 수준은 GI과 GII가 각각 76.47% (13/17), 70.59% (12/17) 로 나타났다. 그러나 기존에 알려진 primer와 본 연구에서 제작한 primer를 사용하였을 때 검출된 양성비율이 차이가 나지 않았다. 검출된 노로바이러스를 염기서열 비교를 통한 계통 발생학적 분석 결과, 동천에서 검출된 GI의 경우 1/2/4/5/9/10의 genotype이 GII의 경우 3/4/5/11/13의 genotype으로 분류되었다. 그리고 본 연구에서 검출된 major type 중 GII/4의 경우, 최근 아시아 각국에서 많은 문제를 일으키고 있는 major genotype으로 알려져 노로바이러스에 대한 위험성을 제고하게 하였다. 또한, 이러한 결과는 국내의 강, 호수, 하천 등이 비슷한 노로바이러스의 GI,II의 genotype으로 오염되어 있음을 암시하며 수계환경 중 미생물의 질을 개선하기 위한 지속적인 노로바이러스의 모니터링이 요구된다.

Keywords

References

  1. Abbaszadegan, M., Huber, M.S., Gerba, C.P. and Pepper, I.L.: Detection of enteroviruses in groundwater with the PCR. Applied and Environmental Microbiology, 59:1318-1324, 1993.
  2. Anderson, A.D., Garrett, V.D., Sobel, J., Monroe, S.S., Fankhauser, R.L., Schwab K.J., Bresee, J.S., Mead, P.S., Higgins, C., Campana, J. and Glass, R.I.: Outbreak Investigation Team. Multistate Outbreak of Norwalk-like Virus Gastroenteritis Associated with a Common Caterer. American journal of epidemiology, 154:1013-1019, 2001. https://doi.org/10.1093/aje/154.11.1013
  3. Ando, T., Noel, J.S. and Fankhauser, R.L.: Genetic classification of "Norwalk-like viruses". Journal of Infectious Diseases, 181 (Suppl. 2): S336-S348. 2000. https://doi.org/10.1086/315589
  4. Atmar, R.L., Neill, F.H., Romalde, J.L., Le Guyader, F., Woodley, C.M., Metcalf, T.G. and Estes, M.K.: Detection of Norwalk virus and hepatitis A viruses in shellfish tissues with the PCR. Applied and Environmental Microbiology, 61: 3014-30189, 2005
  5. Belliot, G., Sosnovtsev, S.V., Mitra, T., Hammer, C., Garfield, M. and Green, K.Y.: In vitro proteolytic processing of the MD145 norovirus ORF1 nonstructural polyprotein yields stable precursors and products similar to those detected in Calicivirus-infected cells. Journal of Virology, 77:10957-10974, 2003. https://doi.org/10.1128/JVI.77.20.10957-10974.2003
  6. Bertolotti-Ciarlet, A., Crawford, S.E., Hutson, A.M. and Estes, M.K.: The 3' end of Norwalk virus mRNA contains determinants that regulate the expression and stability of the capsid protein VP1: a novel function for the VP2 protein. Journal of Virology, 77:11603-11615, 2003. https://doi.org/10.1128/JVI.77.21.11603-11615.2003
  7. Chakravarty, S., Hutson, A.M., Estes, M.K. and Prasad B.V.V.: Evolutionary trace residues in Noroviruses: importance in receptor binding, antigenicity, virion assembly, and strain diversity. Journal of Virology, 79:554-568, 2005 https://doi.org/10.1128/JVI.79.1.554-568.2005
  8. Dolin, R., Blacklow, N.R., DuPont, H., Buscho, R.F., Wyatt, R.G., Kasel, J.A., Hornick, R. and Chanock, R.M.: Biological properties of Norwalk agent of acute infectious nonbacterial gastroenteritis. Proceedings of the Society for Experimental Biology and Medicine, 140(2):578-583, 1972.
  9. Gong, Y.W., Oh, B.Y., Kim, H.Y., Lee, M.Y., Kim, Y.H., Go, J.M. Lee, J.M., Jeong, H.S. and Cheon, D.S.: Molecular Epidemiology Investigation of Norovirus infections in Incheon City, Korea, from 2005 to 2007. Journal of Bacteriology and Virology, 38:249-257, 2008. https://doi.org/10.4167/jbv.2008.38.4.249
  10. Ho, E.C., Cheng, P.K., Lau, A.W. and Lim, W.W.: Atypical norovius epidemic in Hong Knog during summer of 2006 caused by a new genogroup II/4 variant. Journal of Clinical Microbiology, 45:2205-2211, 2007. https://doi.org/10.1128/JCM.02489-06
  11. Jaykus, L.A., De Leon, R. and Sobsey, M.D.: A virion concentration method for detection of human enteric viruses in oysters by PCR and oligoprobe hybridization. Applied and Environmental Microbiology, 62: 2074-2080, 1996.
  12. Jiang, X., Graham, D.Y., Wang, K. and Estes, M.K.: Norwalk virus genome cloning and characterization. Science 250:1580-1583, 1990. https://doi.org/10.1126/science.2177224
  13. Jiang, X., Wang, M., Wang, K. and Estes, M.K.: Sequence and genomic organization of Norwalk virus. Virology 195:51-61, 1993. https://doi.org/10.1006/viro.1993.1345
  14. Kageyama, T., Kojima, S., Shinohara, M., Uchida, K., Fukushi, S., Hoshino, F.B., Takeda, N. and Katayama, K.: Broadly reactive and highly sensitive assay for Norwalk-like viruses Based on Real-time quantitative Reverse Transcription-PCR. Journal of Clinical Microbiology, 41:1548-1557, 2003. https://doi.org/10.1128/JCM.41.4.1548-1557.2003
  15. Kageyama, T., Shinohara, M., Uchida, K., Fukushi, S., Hoshino, F.B., Kojima, S., Takai R., Oka, T., Takeda, N. and Katayama, K.: Coexistence of multiple genotypes, including newly identified genotypes, in outbreaks of gastroenteritis due to Norovirus in Japan. Journal of Clinical Microbiology, 42:2988-2995, 2004. https://doi.org/10.1128/JCM.42.7.2988-2995.2004
  16. Kapikian, A.Z., Wyatt, R.G., Dolin, R., Thornhill, T.S., Kalica, A.R. and Chanock, R.M.: Visualization by immune electron microscopy of a 27 nm particle associated with acute infectious nonbacterial gastroenteritis. Journal of Virology, 10:1075-1081, 1972.
  17. Katayama, H., Simassaki, A., and Ohagaki, S.: Development of a virus concentration method and its application to detection of enterovirus and Norwalk virus from coastal seawater. Applied and Environmental Microbiology, 68:1033-1039, 2002. https://doi.org/10.1128/AEM.68.3.1033-1039.2002
  18. Katayama, K., Shirato-Horikoshi, H., Kojima, S., Kageyama, T., Oka, T., Hoshino, F.B., Fukushi, S., Shinohara, M., Uchida, K., Suzuki, Y., Gojobori, T. and Takeda, N.: Phylogenetic analysis of the complete genome of 18 Norwalk-like viruses. Virology 299:225-239, 2002. https://doi.org/10.1006/viro.2002.1568
  19. Kim, S.H., Cheon, D.S., Kim, J.H., Lee, D.H., Jheong, W.H., Heo, Y.J., Chung, H.M., Jee, Y. and Lee, J.S.: Outbreaks of gastroenteritis that occurred during school excursions in Korea were associated with several waterborne strains of norovirus. Journal of Clinical Microbiology, 43(9):4836-4839, 2005. https://doi.org/10.1128/JCM.43.9.4836-4839.2005
  20. Kojima, S., Kageyama, T., Fukushi, S., Hoshino, F,B., Shinohara, M., Uchida, K., Natori, K., Takeda, N. and Katayama, K.: Genogroup-specific PCR primers for detection of Norwalk-like viruses. Journal of virological methods, 100:107-114, 2002. https://doi.org/10.1016/S0166-0934(01)00404-9
  21. Lee, C. and Kim, S.J.: The genetic diversity of human noroviruses detected in river water in Korea. Water research, 42(17):4477-4484, 2008 https://doi.org/10.1016/j.watres.2008.08.003
  22. Mullendore, J.L., Sobey, M.D. and Shieh, Y.C.: Improved method for the recovery of hepatitis A virus from oyster. Journal of Virology Methods 94:25-35, 2001. https://doi.org/10.1016/S0166-0934(01)00263-4
  23. Lewis, G.D. and Meticalf, T.G.: Polyethylene Glycol Precipitation for Recovery of Pathogenic Viruses, Including Hepatitis A Virus and Human Rotavirus, From Oyster, water and Sediment Samples. Applied and Environmental Microbiology, 54:1983-1988, 1988.
  24. Nishida, T., Nishio, O., Kato, M., Chuma, T., Kato, H., Iwata, H. and Kimura, H.: Genotyping and quantitation of noroviruses in oysters from two distinct sea areas in Japan. Microbiology and Immunology, 51(2):177-184, 2007. https://doi.org/10.1111/j.1348-0421.2007.tb03899.x
  25. Okada, M., Ogawa, T., Kaiho, I. and Shinozaki, K.: Genetic analysis of norovirueses in Chiba Prefecture, Japan, between 1999 and 2004. Journal of Clinical Microbiology, 43:4391-4401, 2005. https://doi.org/10.1128/JCM.43.9.4391-4401.2005
  26. Park, Y.B. and Ko, G.P.: Development of immunomagnetic separation and receptor-mediated methods for noroviruses. ASM meeting, 2007.
  27. Phan, T.G., Khamrin, P., Akiyama, M., Yagyu, F., Okitsu, S., Maneekarn, N., Nishio, O. and Ushijima, H.: Detection and genetic characterization of norovirus in oysters from China and Japan. Clinical Laboratory, 53:405-412, 2007.
  28. Richards, G.P., Watson, M.A., Fankhauser, R.L. and Monroe, S.S.: Genogroup I and II Norovirues Detected in Stool Samples by Real-Time Reverse Transcription-PCR Using Highly Degenerate Universal Primer. Applied and Environmental Microbiology, 70:7179-7184, 2004. https://doi.org/10.1128/AEM.70.12.7179-7184.2004
  29. Center for Disease Control and Prevention. Norovirus activity-United States. Morbidiy and Mortality Weekly Report, 52:41-45, 2002.

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