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. 1984 Feb;19(2):267–272. doi: 10.1128/jcm.19.2.267-272.1984

Enzyme immunoassay for rabies antibody in hybridoma culture fluids and its application to differentiation of street and laboratory strains of rabies virus.

J S Smith, J W Sumner, L F Roumillat
PMCID: PMC271035  PMID: 6365963

Abstract

A rapid and sensitive enzyme immunoassay is described for detecting rabies antibody in hybridoma culture fluids. Glass fiber filter disks were used to immobilize gamma-irradiated mouse neuroblastoma cells infected with street or laboratory strains of rabies virus. Bound rabies-specific antibody was detected by reaction with horseradish peroxidase-labeled goat anti-mouse immunoglobulin G. The assay was performed in a 96-well filtration device developed by Cleveland et al. (J. Clin. Microbiol. 15:402-407, 1982) for the typing of herpes simplex viruses. When partially disrupted cells were used, both internal and external viral antigens were available for reaction. The procedure is rapid (less than 4 h for completion) and requires only small amounts of fluid, and the gamma-irradiated antigen is noninfectious. When the procedure was used to screen 145 fluids from rabies-immune spleen-myeloma cell fusions, 132 were positive for rabies antibody. Other commonly used assays for the detection of rabies-specific antibody were less sensitive. Simultaneous analyses of many hybridoma fluids against a battery of street and laboratory strains of rabies virus are possible and allow rapid selection of useful monoclones.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abelseth M. K. An Attenuated Rabies Vaccine for Domestic Animals Produced in Tissue Culture. Can Vet J. 1964 Nov;5(11):279–286. [PMC free article] [PubMed] [Google Scholar]
  2. Atanasiu P., Savy V., Perrin P. Epreuve immunoenzymatique pour la détection rapide des anticorps antirabiques. Ann Microbiol (Paris) 1977 May-Jun;128A(4):489–498. [PubMed] [Google Scholar]
  3. Charlton K. M., Casey G. A., Boucher D. W., Wiktor T. J. Antigenic variants of rabies virus. Comp Immunol Microbiol Infect Dis. 1982;5(1-3):113–115. doi: 10.1016/0147-9571(82)90023-6. [DOI] [PubMed] [Google Scholar]
  4. Cleveland P. H., Richman D. D., Oxman M. N., Worthen D. M. Rapid serological technique for typing herpes simplex viruses. J Clin Microbiol. 1982 Mar;15(3):402–407. doi: 10.1128/jcm.15.3.402-407.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Elliott L. H., McCormick J. B., Johnson K. M. Inactivation of Lassa, Marburg, and Ebola viruses by gamma irradiation. J Clin Microbiol. 1982 Oct;16(4):704–708. doi: 10.1128/jcm.16.4.704-708.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Flamand A., Wiktor T. J., Koprowski H. Use of hybridoma monoclonal antibodies in the detection of antigenic differences between rabies and rabies-related virus proteins. II. The glycoprotein. J Gen Virol. 1980 May;48(1):105–109. doi: 10.1099/0022-1317-48-1-105. [DOI] [PubMed] [Google Scholar]
  7. Gamble W. C., Chappell W. A., George E. H. Inactivation of rabies diagnostic reagents by gamma radiation. J Clin Microbiol. 1980 Nov;12(5):676–678. doi: 10.1128/jcm.12.5.676-678.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hummeler K., Koprowski H., Wiktor T. J. Structure and development of rabies virus in tissue culture. J Virol. 1967 Feb;1(1):152–170. doi: 10.1128/jvi.1.1.152-170.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Iwasaki Y., Gerhard W., Clark H. F. Role of host immune response in the development of either encephalitic or paralytic disease after experimental rabies infection in mice. Infect Immun. 1977 Oct;18(1):220–225. doi: 10.1128/iai.18.1.220-225.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. KISSLING R. E. Growth of rabies virus in non-nervous tissue culture. Proc Soc Exp Biol Med. 1958 Jun;98(2):223–225. doi: 10.3181/00379727-98-23997. [DOI] [PubMed] [Google Scholar]
  11. KOPROWSKI H. Biological modification of rabies virus as a result of its adaptation to chicks and developing chick embryos. Bull World Health Organ. 1954;10(5):709–724. [PMC free article] [PubMed] [Google Scholar]
  12. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  13. Lodmell D. L., Arai Y. T., Ewalt L. C. Influence of cell type and virus upon lysis of rabies virus-infected cells by antibody and complement. Arch Virol. 1981;70(2):147–155. doi: 10.1007/BF01315008. [DOI] [PubMed] [Google Scholar]
  14. Schneider L. G., Dietzschold B., Dierks R. E., Matthaeus W., Enzmann P. J., Strohmaier K. Rabies group-specific ribonucleoprotein antigen and a test system for grouping and typing of rhabdoviruses. J Virol. 1973 May;11(5):748–755. doi: 10.1128/jvi.11.5.748-755.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sedwick W. D., Wiktor T. J. Reproducible plaquing system for rabies, lymphocytic choriomeningitis,k and other ribonucleic acid viruses in BHK-21-13S agarose suspensions. J Virol. 1967 Dec;1(6):1224–1226. doi: 10.1128/jvi.1.6.1224-1226.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Smith J. S., Yager P. A., Baer G. M. A rapid reproducible test for determining rabies neutralizing antibody. Bull World Health Organ. 1973 May;48(5):535–541. [PMC free article] [PubMed] [Google Scholar]
  17. Thraenhart O., Kuwert E. K. Enzyme immunoassay for demonstration of rabies-virus antibodies after immunisation. Lancet. 1977 Aug 20;2(8034):399–400. doi: 10.1016/s0140-6736(77)90321-x. [DOI] [PubMed] [Google Scholar]
  18. Wiktor T. J., Clark H. F. Chronic rabies virus infection of cell cultures. Infect Immun. 1972 Dec;6(6):988–995. doi: 10.1128/iai.6.6.988-995.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wiktor T. J., György E., Schlumberger D., Sokol F., Koprowski H. Antigenic properties of rabies virus components. J Immunol. 1973 Jan;110(1):269–276. [PubMed] [Google Scholar]

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