1887

Abstract

Summary

A monoclonal radioimmunoassay (RIA) was developed for detection of dengue virus in infected cell culture fluids and blood samples from dengue patients. Antibodies used to construct the RIA were selected on the basis of high binding avidity, the demonstration of synergism in competitive binding assays and empirical trials with different antibody combinations. Optimal binding of all four dengue virus serotypes was achieved by use of a flavivirus group-reactive and a dengue virus complex-reactive antibody attached to a solid-phase support and a single flavivirus subgroup-reactive antibody as radiolabelled probe. A ‘simultaneous sandwich’ format and prolonged (18 h) incubation at 37 °C yielded optimal results. The limit of sensitivity of the RIA for detection of dengue type 2 virus was 2.7 log mosquito 50% infectious doses (MID). The assay was tenfold more sensitive for dengue type 2 than for dengue types 1 and 3 viruses and 100-fold more sensitive than for dengue type 4 virus. Specificity, assessed using over 500 disease control human sera, was increased by addition of monoclonal anti-tetanus blocking antibodies, resulting in a false positive rate of only 0.2%. Heterologous dengue virus antibodies were shown to inhibit the RIA in assays performed with artificial immune complexes. Acute phase human sera containing 10 to 10 MID but no detectable antigen by RIA, were also shown to inhibit binding of the homologous dengue virus serotype; this effect was attributed to heterologous antibody from a prior infection. Among 116 viraemic sera from dengue patients, the RIA was positive in 43 to 47% of patients with dengue type 1, 2 or 3 infections but in only 10% of the dengue type 4 cases. Virus was more frequently detected in cases of primary infection (54%) than in cases of superinfection (16%). Despite the limitations imposed by immunological interference, the antigen capture RIA appears useful as a rapid diagnostic technique for dengue surveillance.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-67-4-639
1986-04-01
2022-05-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/67/4/JV0670040639.html?itemId=/content/journal/jgv/10.1099/0022-1317-67-4-639&mimeType=html&fmt=ahah

References

  1. Brandt W. E., Cardiff R. D., Russell P. K. 1970; Dengue virions and antigens in brain and serum of infected mice. Journal of Virology 6:500–506
    [Google Scholar]
  2. Clarke D. H., Casals I. 1958; Techniques for hemagglutination and hemagglutination-inhibition with arthropod-borne viruses. American Journal of Tropical Medicine and Hygiene 7:561–573
    [Google Scholar]
  3. Ey P. L., Prowse S. J., Jenkin C. R. 1978; Isolation of pure IgGl, IgG2a, and IgG2b immunoglobulins from mouse serum using Protein A-Sepharose. Immunochemistry 15:429–436
    [Google Scholar]
  4. Fraker P. J., Speck J. C. 1978; Protein and cell membrane iodinations with a sparingly soluble chloramide, l, 3, 4, 6-tetrachloro-3a, 6a-diphenyl glycoluril. Biochemical and Biophysical Research Communications 80:849–857
    [Google Scholar]
  5. Frankel M. E., Gerhard W. 1979; The rapid determination of binding constants for antiviral antibodies by a radioimmunoassay. An analysis of the interaction between hybridoma proteins and influenza virus. Molecular Immunology 16:101–105
    [Google Scholar]
  6. Gentry M. K., Henchal E. A., Mccown J. M., Brandt W. E., Dalrymple J. M. 1982; Identification of distinct determinants on dengue-2 virus using monoclonal antibodies. American Journal of Tropical Medicine and Hygiene 31:548–555
    [Google Scholar]
  7. Gubler D. J., Suharyono W., Tan R., Abidin M., Sie A. 1981; Viraemia in patients with naturally acquired dengue infection. Bulletin of the World Health Organization 59:623–630
    [Google Scholar]
  8. Halstead S. B. 1980; Immunopathological parameters of toga virus disease syndromes. In The Togaviruses : Biology, Structure, Replication pp. 107–174 Edited by Schlesinger R. W. New York & London: Academic Press;
    [Google Scholar]
  9. Halstead S. B., Rojanasuphot S., Sangkawibha N. 1983; Original antigenic sin in dengue. American Journal of Tropical Medicine and Hygiene 32:154–156
    [Google Scholar]
  10. Heinz F. X., Mandle C., Berger R., Tuma W., Kunz C. 1984; Antibody-induced conformational changes result in enhanced avidity of antibodies to different antigenic sites on the tick-borne encephalitis virus glycoprotein. Virology 133:25–34
    [Google Scholar]
  11. Henchal E. A., Mccown J. M., Burke D. S., Sequin M. C., Brandt W. E. 1985; Epitopic analysis of antigenic determinants on the surface of dengue-2 virions using monoclonal antibodies. American Journal of Tropical Medicine and Hygiene 34:162–169
    [Google Scholar]
  12. Monath T. P., Hill L. H., Brown N. V., Cropp C. B., Schlesinger J. J., Saluzzo J. F., Wands I. R. 1986; Sensitive and specific monoclonal immunoassay for detecting yellow fever virus in laboratory and clinical specimens. Journal of Clinical Microbiology (in press)
    [Google Scholar]
  13. Rosen L., Gubler D. J. 1974; The use of mosquitoes to detect and propagate dengue viruses. American Journal of Tropical Medicine and Hygiene 23:1153–1160
    [Google Scholar]
  14. Sangkawibha N., Rojanasuphot S., Ahandrik S., Viriyapongse S., Jatanasen S., Salitul V., Phanthuma-Chinda B., Halstead S. B. 1984; Risk factors in dengue shock syndrome: a prospective epidemiologic study in Rayong, Thailand. 1. The 1980 outbreak. American Journal of Epidemiology 120:653–669
    [Google Scholar]
  15. Schlesinger J. J., Walsh E. E., Brandiss M. W. 1984; Analysis of 17D yellow fever virus envelope protein epitopes using monoclonal antibodies. Journal of General Virology 65:1637–1644
    [Google Scholar]
  16. Theofilopoulos A. N., Wilson C. B., Dixon F. J. 1976; The Raji cell radioimmune assay for detecting immune complexes in human sera. Journal of Clinical Investigation 57:169–182
    [Google Scholar]
  17. Trent D. W., Grant J. A. 1980; A comparison of New World alphaviruses in the western equine encephalomyelitis complex by immunochemical and oligonucleotide fingerprint techniques. Journal of General Virology 47:261–282
    [Google Scholar]
  18. Wands J. R., Carlson R. I., Schoemaker H., Isselbacher K. J., Zurawski V. R. Jr 1981; Immunodiagnosis of hepatitis B with high-affinity IgM monoclonal antibodies. Proceedings of the National Academy of Sciences, U,. S,. A 78:1214–1218
    [Google Scholar]
  19. Zurawski V. R. Jr, Hurrell J. G. R., Lathan W. C., Black P. H., Haber E. 1980; Monoclonal antibodies to tetanus toxin produced by clones of murine hybridomas. Federation Proceedings 394922
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-67-4-639
Loading
/content/journal/jgv/10.1099/0022-1317-67-4-639
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error