VP4 and VP7 typing using monoclonal antibodies

  • B. S. Coulson
Part of the Archives of Virology book series (ARCHIVES SUPPL, volume 12)


Both rotavirus outer capsid proteins, VP4 and VP7, elicit neutralizing antibodies. Neutralizing mouse monoclonal antibodies (N-MAbs) to VP7 are easily derived and have been used widely and successfully to serotype both stool-derived and culture-adapted rotaviruses by enzyme immunoassay (EIA). Generally, approximately 70% of rotaviruses in stool samples are typable by VP7 EIA, an inexpensive and practical method. Variations in antigenic regions between strains within human rotavirus serotypes 1, 2, 4, and 9 have been recorded. These have been termed monotypes because they are detected with N-MAbs. The molecular basis for monotypes has been determined by mapping mutations selected in N-MAb-resistant antigenic variants, and by sequence analysis of the gene encoding VP7 in newly recognized monotypes. Antigenic regions A, B and C in VP7 are involved. In order to detect all members of a particular VP7 serotype, it is necessary to type with a panel of N-MAbs specific for that serotype.

N-MAbs to VP4 of human rotavirus are difficult to raise and few have proven suitable for VP4 serotyping by EIA. The specificity of the assay for each P type is highest when the VP7 serotype specificity of the capture antiserum is matched to the G type of the rotavirus in the test sample. The VP4 EIA gives similar typing rates to the VP7 typing EIA. N-MAbs directed to VP8*, the smaller subunit of VP4 generated by proteolytic cleavage, are more likely to show serotype specificity. Some N-MAbs that select mutations in the putative fusion region of VP5*, the larger subunit of VP4, show cross-reactivity with extracts of normal, uninfected MA 104 cells and with fetal bovine serum. These N-MAbs also give elevated EIA OD readings with rotavirus-positive, but previously non-reactive fecal samples which have been frozen and thawed repeatedly. Overall, VP8*-reactive N-MAbs appear most suitable for VP4 typing by EIA.


Neutralization Epitope Antigenic Region Outer Capsid Protein Elicit Neutralize Antibody Typing Monoclonal Antibody 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Chen D, Estes MK, Ramig RF (1992) Specific interactions between rotavirus outer capsid proteins VP4 and VP7 determine expression of a cross-reactive, neutralizing VP4-specific epitope. J Virol 66: 432–439PubMedGoogle Scholar
  2. 2.
    Coulson BS (1987) Variation in neutralization epitopes of human rotaviruses in relation to genomic RNA polymorphism. Virology 159: 209–216PubMedCrossRefGoogle Scholar
  3. 3.
    Coulson BS (1993) Typing of human rotavirus VP4 by an enzyme immunoassay using monoclonal antibodies. J Clin Microbiol 31: 1–8PubMedGoogle Scholar
  4. 4.
    Coulson BS, Fowler KJ, Bishop RF, Cotton RGH (1985) Neutralizing monoclonal antibodies to human rotavirus and indications of antigenic drift among strains from neonates. J Virol 54: 14–20PubMedGoogle Scholar
  5. 5.
    Coulson BS, Kirkwood C (1991) Relation of VP7 amino acid sequence to monoclonal antibody neutralization of rotavirus and rotavirus monotypes. J Virol 65: 5968–5974PubMedGoogle Scholar
  6. 6.
    Coulson BS, Tursi JM, McAdam WJ, Bishop RF (1986) Derivation of neutralizing monoclonal antibodies to human rotaviruses and evidence that an immunodominant neutralization site is shared between serotypes 1 and 3. Virology 154: 302–312PubMedCrossRefGoogle Scholar
  7. 7.
    Coulson BS, Unicomb LE, Pitson GA, Bishop RF (1987) Simple and specific enzyme immunoassay using monoclonal antibodies for serotyping human rotaviruses. J Clin Microbiol 25: 509–515PubMedGoogle Scholar
  8. 8.
    Estes MK (1996) Rotaviruses and their replication. In: Fields BN, Knipe DM, Howley PM, Chanock RM, Melnick JL, Monath TP, Roizman B, Strauss S (eds) Virology. Lippincott-Raven Publishers, Philadelphia, pp 1625–1655Google Scholar
  9. 9.
    Gentsch JR, Glass RI, Woods P, Gouvea V, Gorziglia M, Flores J, Das BK, Bhan MK (1992) Identification of group A rotavirus gene 4 types by polymerase chain reaction. J Clin Microbiol 30: 1365–1373PubMedGoogle Scholar
  10. 10.
    Gerna G. Sarasini A, Matteo A, Parea M, Orsolini P, Battaglia M (1988) Identification of two subtypes of serotype 4 human rotavirus by using VP7-specific neutralizing monoclonal antibodies. J Clin Microbiol. 26: 1388–1392PubMedGoogle Scholar
  11. 11.
    Gorziglia M, Larralde G, Kapikian AZ, Chanock RM (1990) Antigenic relationships among human rotaviruses as determined by outer capsid protein VP4. Proc Natl Acad Sci USA 87: 7155–7159PubMedCrossRefGoogle Scholar
  12. 12.
    Hoshino Y, Sereno MM, Midthun K, Flores J, Kapikian AZ, Chanock RM (1985) Independent segregation of two antigenic specifications (VP3 and VP7) involved in neutralization of rotavirus infectivity. Proc Natl Acad Sci USA 82: 8701–8704PubMedCrossRefGoogle Scholar
  13. 13.
    Kirkwood CD, Bishop RF, Coulson BS (1996) Human rotavirus VP4 contains strain-specific and cross-reactive neutralization sites. Arch Virol 141: 587–600PubMedCrossRefGoogle Scholar
  14. 14.
    Kobayashi N, Taniguchi K, Urasawa S (1990) Identification of operationally overlapping and independent cross-reactive neutralization regions on human rotavirus VP4. J Gen Virol 71: 2615–2623PubMedCrossRefGoogle Scholar
  15. 15.
    Kobayashi N, Taniguchi K, Urasawa T, Urasawa S (1991) Preparation and characterization of a neutralizing monoclonal antibody directed to VP4 of rotavirus strain K8 which has unique VP4 neutralization epitopes. Arch Virol 121: 153–162PubMedCrossRefGoogle Scholar
  16. 16.
    Larralde G, Gorziglia M (1992) Distribution of conserved and specific epitopes on the VP8 subunit of rotavirus VP4. J Virol 65: 3213–3218Google Scholar
  17. 17.
    Larralde G, Li B, Kapikian AZ, Gorziglia M (1991) Serotype-specific epitope(s) present on the VP8 subunit of rotavirus VP4 protein. J Virol 65: 3213–3218PubMedGoogle Scholar
  18. 18.
    Padilla-Noriega L, Dunn SJ, Lopez S, Greenberg HB, Arias CF (1995) Identification of two independent neutralization domains on the VP4 trypsin cleavage products VP5* and VP8* of human rotavirus ST-3. Virology 206: 148–154PubMedCrossRefGoogle Scholar
  19. 19.
    Padilla-Noriega L, Werner-Eckert R, Mackow ER, Gorziglia M, Larralde G, Taniguchi K, Greenberg HB (1993) Serologic analysis of human rotavirus serotypes P1A and P2 by using monoclonal antibodies. J Clin Microbiol 31: 622–628PubMedGoogle Scholar
  20. 20.
    Steele AD, Van Niekerk MC, Mphahlele MJ (1995) Geographic distribution of human rotavirus VP4 genotypes and VP7 serotypes in five South African regions. J Clin Microbiol 33: 1516–1519PubMedGoogle Scholar
  21. 21.
    Taniguchi K, Maloy WL, Nishikawa K, Green KY, Hoshino Y, Urasawa S, Kapikian AZ, Chanock RM, Gorziglia M (1988) Identification of cross-reactive and serotype 2-specific neutralization epitopes on VP3 of human rotavirus. J Virol 62: 2421–2426PubMedGoogle Scholar
  22. 22.
    Taniguchi K, Morita Y, Urasawa S (1987) Cross-reactive neutralization epitopes on VP3 of human rotavirus: analysis with monoclonal antibodies and antigenic variants. J Virol 61: 1726–1730PubMedGoogle Scholar
  23. 23.
    Taniguchi K, Urasawa T, Morita Y, Greenberg HB, Urasawa S (1987) Direct serotyping of human rotavirus in stools by an enzyme-linked immunosorbent assay using serotype 1-, 2-, 3-, 4-specific monoclonal antibodies to VP7. J Infect Dis 155: 1159–1165PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 1996

Authors and Affiliations

  • B. S. Coulson
    • 1
  1. 1.Department of MicrobiologyThe University of MelbourneParkvilleAustralia

Personalised recommendations