Archives of Virology

, Volume 151, Issue 3, pp 551–561 | Cite as

Development of an antigen ELISA to detect sapovirus in clinical stool specimens

  • G. S. Hansman
  • R. Guntapong
  • Y. Pongsuwanna
  • K. Natori
  • K. Katayama
  • N. Takeda


Human sapovirus (SaV) strains are etiological agents of mild and/or acute gastroenteritis in children and adults. In this study, we describe the development of a novel antigen enzyme-linked immunosorbent assay (ELISA) detection system that was based on hyperimmune rabbit and guinea pig antisera raised against SaV genogroup I (GI) virus-like particles. The ELISA had 100% specificity, and sensitivities of 60% and 25% when compared to single-round PCR and nested PCR, respectively. Our results have shown the ELISA was useful in detecting SaV GI antigens in clinical stool specimens collected two days after the onset of illness.


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  1. Belliot, G, Noel, JS, Li, JF, Seto, Y, Humphrey, CD, Ando, T, Glass, RI, Monroe, SS 2001Characterization of capsid genes, expressed in the baculovirus system, of three new genetically distinct strains of “Norwalk-like viruses”.J Clin Microbiol3942884295CrossRefPubMedGoogle Scholar
  2. Chiba, S, Sakuma, Y, Kogasaka, R, Akihara, M, Horino, K, Nakao, T, Fukui, S 1979An outbreak of gastroenteritis associated with calicivirus in an infant home.J Med Virol4249254PubMedGoogle Scholar
  3. Chiba, S, Sakuma, Y, Kogasaka, R, Akihara, M, Terashima, H, Horino, K, Nakao, T 1980Fecal shedding of virus in relation to the days of illness in infantile gastroenteritis due to calicivirus.J Infect Dis142247249PubMedGoogle Scholar
  4. Farkas, T, Zhong, WM, Jing, Y, Huang, PW, Espinosa, SM, Martinez, N, Morrow, AL, Ruiz-Palacios, GM, Pickering, LK, Jiang, X 2004Genetic diversity among sapoviruses.Arch Virol14913091323CrossRefPubMedGoogle Scholar
  5. Green, J, Henshilwood, K, Gallimore, CI, Brown, DW, Lees, DN 1998A nested reverse transcriptase PCR assay for detection of small round-structured viruses in environmentally contaminated molluscan shellfish.Appl Environ Microbiol64858863PubMedGoogle Scholar
  6. Guntapong, R, Hansman, G, Oka, T, Ogawa, S, Kageyama, T, Pongsuwanna, Y, Katayama, K 2004Norovirus and sapovirus infections in Thailand.J Infect Dis57276278Google Scholar
  7. Hansman, G, Doan, LP, Kguyen, T, Okitsu, S, Katayama, K, Ogawa, S, Natori, K, Takeda, N, Kato, Y, Nishio, O, Noda, M, Ushijima, H 2004Detection of norovirus and sapovirus infection among children with gastroenteritis in Ho Chi Minh City, Vietnam.Arch Virol14916731688CrossRefPubMedGoogle Scholar
  8. Hansman, GS, Katayama, K, Maneekarn, N, Peerakome, S, Khamrin, P, Tonusin, S, Okitsu, S, Nishio, O, Takeda, N, Ushijima, H 2004Genetic diversity of norovirus and sapovirus in hospitalized infants with sporadic cases of acute gastroenteritis in Chiang Mai, Thailand.J Clin Microbiol4213051307CrossRefPubMedGoogle Scholar
  9. Hansman, GS, Katayama, K, Oka, T, Natori, K, Takeda, N 2005Mutational study of sapovirus expression in insect cells.Virol J213CrossRefPubMedGoogle Scholar
  10. Hansman, GS, Natori, K, Oka, T, Ogawa, S, Tanaka, K, Nagata, N, Ushijima, H, Takeda, N, Katayama, K 2005Cross-reactivity among sapovirus recombinant capsid proteins.Arch Virol1502136PubMedGoogle Scholar
  11. Hansman, GS, Natori, K, Ushijima, H, Katayama, K, Takeda, N 2005Characterization of polyclonal antibodies raised against sapovirus genogroup five virus-like particles.Arch Virol15014331437PubMedGoogle Scholar
  12. Jiang, X, Cubitt, D, Hu, J, Dai, X, Treanor, J, Matson, DO, Pickering, LK 1995Development of an ELISA to detect MX virus, a human calicivirus in the snow Mountain agent genogroup.J Gen Virol7627392747PubMedGoogle Scholar
  13. Jiang, X, Matson, DO, Ruiz-Palacios, GM, Hu, J, Treanor, J, Pickering, LK 1995Expression, self-assembly, and antigenicity of a snow mountain agent-like calicivirus capsid protein.J Clin Microbiol3314521455PubMedGoogle Scholar
  14. Jiang, X, Huang, PW, Zhong, WM, Farkas, T, Cubitt, DW, Matson, DO 1999Design and evaluation of a primer pair that detects both Norwalk- and Sapporo-like caliciviruses by RT-PCR.J Virol Methods83145154CrossRefPubMedGoogle Scholar
  15. Jiang, X, Wilton, N, Zhong, WM, Farkas, T, Huang, PW, Barrett, E, Guerrero, M, Ruiz-Palacios, G, Green, KY, Green, J, Hale, AD, Estes, MK, Pickering, LK, Matson, DO 2000Diagnosis of human caliciviruses by use of enzyme immunoassays.J Infect Dis181S349S359PubMedGoogle Scholar
  16. Johansson, PJ, Bergentoft, K, Larsson, PA, Magnusson, G, Widell, A, Thorhagen, M, Hedlund, KO 2005A nosocomial sapovirus-associated outbreak of gastroenteritis in adults.Scand J Infect Dis37200204PubMedGoogle Scholar
  17. Katayama, K, Miyoshi, T, Uchino, K, Oka, T, Tanaka, T, Takeda, N, Hansman, GS 2004Novel recombinant sapovirus.Emerg Infect Dis1018741876PubMedGoogle Scholar
  18. Kitamoto, N, Tanaka, T, Natori, K, Takeda, N, Nakata, S, Jiang, X, Estes, MK 2002Cross-reactivity among several recombinant calicivirus virus-like particles (VLPs) with monoclonal antibodies obtained from mice immunized orally with one type of VLP.J Clin Microbiol4024592465CrossRefPubMedGoogle Scholar
  19. Kobayashi, S, Sakae, K, Natori, K, Takeda, N, Miyamura, T, Suzuki, Y 2000Serotype-specific antigen ELISA for detection of Chiba virus in stools.J Med Virol62233238CrossRefPubMedGoogle Scholar
  20. Kobayashi, S, Sakae, K, Suzuki, Y, Ishiko, H, Kamata, K, Suzuki, K, Natori, K, Miyamura, T, Takeda, N 2000Expression of recombinant capsid proteins of chitta virus, a genogroup II Norwalk virus, and development of an ELISA to detect the viral antigen.Microbiol Immunol44687693PubMedGoogle Scholar
  21. Kobayashi, S, Sakae, K, Suzuki, Y, Shinozaki, K, Okada, M, Ishiko, H, Kamata, K, Suzuki, K, Natori, K, Miyamura, T, Takeda, N 2000Molecular cloning, expression, and antigenicity of Seto virus belonging to genogroup I Norwalk-like viruses.J Clin Microbiol3834923494PubMedGoogle Scholar
  22. Kojima, S, Kageyama, T, Fukushi, S, Hoshino, FB, Shinohara, M, Uchida, K, Natori, K, Takeda, N, Katayama, K 2002Genogroup-specific PCR primers for detection of Norwalk-like viruses.J Virol Methods100107114CrossRefPubMedGoogle Scholar
  23. Matson, DO, Estes, MK, Glass, RI, Bartlett, AV, Penaranda, M, Calomeni, E, Tanaka, T, Nakata, S, Chiba, S 1989Human calicivirus-associated diarrhea in children attending day care centers.J Infect Dis1597178PubMedGoogle Scholar
  24. McIver, CJ, Hansman, G, White, P, Doultree, JC, Catton, M, Rawlinson, WD 2001Diagnosis of enteric pathogens in children with gastroenteritis.Pathology33353358PubMedGoogle Scholar
  25. Nakata, S, Chiba, S, Terashima, H, Sakuma, Y, Kogasaka, R, Nakao, T 1983Microtiter solid-phase radioimmunoassay for detection of human calicivirus in stools.J Clin Microbiol17198201PubMedGoogle Scholar
  26. Nakata, S, Chiba, S, Terashima, H, Nakao, T 1985Prevalence of antibody to human calicivirus in Japan and Southeast Asia determined by radioimmunoassay.J Clin Microbiol22519521PubMedGoogle Scholar
  27. Nakata, S, Chiba, S, Terashima, H, Yokoyama, T, Nakao, T 1985Humoral immunity in infants with gastroenteritis caused by human calicivirus.J Infect Dis152274279PubMedGoogle Scholar
  28. Nakata, S, Estes, MK, Chiba, S 1988Detection of human calicivirus antigen and antibody by enzyme-linked immunosorbent assays.J Clin Microbiol2620012005PubMedGoogle Scholar
  29. Nakata, S, Honma, S, Numata, K, Kogawa, K, Ukae, S, Adachi, N, Jiang, X, Estes, MK, Gatheru, Z, Tukei, PM, Chiba, S 1998Prevalence of human calicivirus infections in Kenya as determined by enzyme immunoassays for three genogroups of the virus.J Clin Microbiol3631603163PubMedGoogle Scholar
  30. Okada, M, Shinozaki, K, Ogawa, T, Kaiho, I 2002Molecular epidemiology and phylogenetic analysis of Sapporo-like viruses.Arch Virol14714451451CrossRefPubMedGoogle Scholar
  31. Rabenau, HF, Sturmer, M, Buxbaum, S, Walczok, A, Preiser, W, Doerr, HW 2003Laboratory diagnosis of norovirus: which method is the best?Intervirology46232238CrossRefPubMedGoogle Scholar
  32. Sakuma, Y, Chiba, S, Kogasaka, R, Terashima, H, Nakamura, S, Horino, K, Nakao, T 1981Prevalence of antibody to human calicivirus in general population of northern Japan.J Med Virol7221225PubMedGoogle Scholar
  33. Vinje, J, Deijl, H, van der Heide, R, Lewis, D, Hedlund, KO, Svensson, L, Koopmans, MP 2000Molecular detection and epidemiology of Sapporo-like viruses.J Clin Microbiol38530536PubMedGoogle Scholar

Copyright information

© Springer-Verlag/Wien 2005

Authors and Affiliations

  • G. S. Hansman
    • 1
  • R. Guntapong
    • 2
  • Y. Pongsuwanna
    • 2
  • K. Natori
    • 1
  • K. Katayama
    • 1
  • N. Takeda
    • 1
  1. 1.Department of Virology IINational Institute of Infectious DiseasesTokyoJapan
  2. 2.Department of Medical SciencesNational Institute of Health, Ministry of Public HealthThailand

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