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. 1997 Jul;4(4):452–457. doi: 10.1128/cdli.4.4.452-457.1997

Evaluation of a dipstick enzyme-linked immunosorbent assay for detection of antibodies to dengue virus.

S J Wu 1, B Hanson 1, H Paxton 1, A Nisalak 1, D W Vaughn 1, C Rossi 1, E A Henchal 1, K R Porter 1, D M Watts 1, C G Hayes 1
PMCID: PMC170549  PMID: 9220163

Abstract

Accurate serological confirmation of dengue (DEN) infection is difficult, because simple reliable assays for the detection of DEN antibodies are not available. To address this problem, a dipstick enzyme-linked immunosorbent assay (ELISA) was evaluated. The dipstick contained dots of serially diluted DEN 2 antigen. To detect immunoglobulin G (IgG), the dipstick was processed through four reaction cuvettes containing test serum, enhancer, enzyme-conjugated anti-human IgG and IgM antibody, and substrate. Total assay time was 45 min. To detect IgM, the serum was passed through a protein G device to remove IgG. The dipstick was then processed as before, except that the incubation times were longer and enzyme-conjugated anti-human IgM was used. The total assay time was 3 h. The dipstick ELISA results were compared with results from microplate ELISA. The IgG dipstick ELISA showed a sensitivity of 95.2% and a specificity of 100% compared to an IgG microplate ELISA with serum samples from 125 individuals living in an area in which DEN is endemic. In tests with 75 serum samples from patients with clinically suspected acute DEN infections, the IgM dipstick ELISA showed a sensitivity of 97.9% and specificity of 100% compared to those of an IgM antibody capture microplate ELISA. These results showed that the dipstick ELISA was a sensitive and specific test for the detection of either DEN IgM or IgG in human serum. The dipstick ELISA was also shown to be useful for detecting seroconversions to DEN IgM or IgG in paired serum samples from 20 patients with virus isolation-confirmed acute DEN infections.

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Selected References

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  1. Ansari M. Z., Shope R. E., Malik S. Evaluation of vero cell lysate antigen for the ELISA of flaviviruses. J Clin Lab Anal. 1993;7(4):230–237. doi: 10.1002/jcla.1860070408. [DOI] [PubMed] [Google Scholar]
  2. Bundo K., Igarashi A. Antibody-capture ELISA for detection of immunoglobulin M antibodies in sera from Japanese encephalitis and dengue hemorrhagic fever patients. J Virol Methods. 1985 May;11(1):15–22. doi: 10.1016/0166-0934(85)90120-x. [DOI] [PubMed] [Google Scholar]
  3. CLARKE D. H., CASALS J. Techniques for hemagglutination and hemagglutination-inhibition with arthropod-borne viruses. Am J Trop Med Hyg. 1958 Sep;7(5):561–573. doi: 10.4269/ajtmh.1958.7.561. [DOI] [PubMed] [Google Scholar]
  4. Cardosa M. J., Baharudin F., Hamid S., Hooi T. P., Nimmanitya S. A nitrocellulose membrane based IgM capture enzyme immunoassay for etiological diagnosis of dengue virus infections. Clin Diagn Virol. 1995 May;3(4):343–350. doi: 10.1016/0928-0197(94)00049-z. [DOI] [PubMed] [Google Scholar]
  5. Cardosa M. J., Tio P. H. Dot enzyme immunoassay: an alternative diagnostic aid for dengue fever and dengue haemorrhagic fever. Bull World Health Organ. 1991;69(6):741–745. [PMC free article] [PubMed] [Google Scholar]
  6. Cardosa M. J., Zuraini I. Comparison of an IgM capture ELISA with a dot enzyme immunoassay for laboratory diagnosis of dengue virus infections. Southeast Asian J Trop Med Public Health. 1991 Sep;22(3):337–340. [PubMed] [Google Scholar]
  7. Chungue E., Marché G., Plichart R., Boutin J. P., Roux J. Comparison of immunoglobulin G enzyme-linked immunosorbent assay (IgG-ELISA) and haemagglutination inhibition (HI) test for the detection of dengue antibodies. Prevalence of dengue IgG-ELISA antibodies in Tahiti. Trans R Soc Trop Med Hyg. 1989 Sep-Oct;83(5):708–711. doi: 10.1016/0035-9203(89)90404-5. [DOI] [PubMed] [Google Scholar]
  8. Halstead S. B. Pathogenesis of dengue: challenges to molecular biology. Science. 1988 Jan 29;239(4839):476–481. doi: 10.1126/science.3277268. [DOI] [PubMed] [Google Scholar]
  9. Halstead S. B. Selective primary health care: strategies for control of disease in the developing world. XI. Dengue. Rev Infect Dis. 1984 Mar-Apr;6(2):251–264. doi: 10.1093/clinids/6.2.251. [DOI] [PubMed] [Google Scholar]
  10. Hayes C. G., Phillips I. A., Callahan J. D., Griebenow W. F., Hyams K. C., Wu S. J., Watts D. M. The epidemiology of dengue virus infection among urban, jungle, and rural populations in the Amazon region of Peru. Am J Trop Med Hyg. 1996 Oct;55(4):459–463. doi: 10.4269/ajtmh.1996.55.459. [DOI] [PubMed] [Google Scholar]
  11. Henchal E. A., Putnak J. R. The dengue viruses. Clin Microbiol Rev. 1990 Oct;3(4):376–396. doi: 10.1128/cmr.3.4.376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Innis B. L., Nisalak A., Nimmannitya S., Kusalerdchariya S., Chongswasdi V., Suntayakorn S., Puttisri P., Hoke C. H. An enzyme-linked immunosorbent assay to characterize dengue infections where dengue and Japanese encephalitis co-circulate. Am J Trop Med Hyg. 1989 Apr;40(4):418–427. doi: 10.4269/ajtmh.1989.40.418. [DOI] [PubMed] [Google Scholar]
  13. Johnson R. B., Jr, Libby R. Separation of immunoglobulin M (IgM) essentially free of IgG from serum for use in systems requiring assay of IgM-type antibodies without interference from rheumatoid factor. J Clin Microbiol. 1980 Sep;12(3):451–454. doi: 10.1128/jcm.12.3.451-454.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kelly D. J., Chan C. T., Paxton H., Thompson K., Howard R., Dasch G. A. Comparative evaluation of a commercial enzyme immunoassay for the detection of human antibody to Rickettsia typhi. Clin Diagn Lab Immunol. 1995 May;2(3):356–360. doi: 10.1128/cdli.2.3.356-360.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Koay A. S., Cheong Y. M. Preliminary evaluation of the INDX DIP-S-TICKS with positive rickettsial samples in Malaysia. Malays J Pathol. 1993 Jun;15(1):69–73. [PubMed] [Google Scholar]
  16. Kuno G., Gómez I., Gubler D. J. An ELISA procedure for the diagnosis of dengue infections. J Virol Methods. 1991 Jun;33(1-2):101–113. doi: 10.1016/0166-0934(91)90011-n. [DOI] [PubMed] [Google Scholar]
  17. Repik P. M., Dalrymple J. M., Brandt W. E., McCown J. M., Russell P. K. RNA fingerprinting as a method for distinguishing dengue 1 virus strains. Am J Trop Med Hyg. 1983 May;32(3):577–589. doi: 10.4269/ajtmh.1983.32.577. [DOI] [PubMed] [Google Scholar]
  18. Sharp T. W., Wallace M. R., Hayes C. G., Sanchez J. L., DeFraites R. F., Arthur R. R., Thornton S. A., Batchelor R. A., Rozmajzl P. J., Hanson R. K. Dengue fever in U.S. troops during Operation Restore Hope, Somalia, 1992-1993. Am J Trop Med Hyg. 1995 Jul;53(1):89–94. [PubMed] [Google Scholar]
  19. Songco R. S., Hayes C. G., Leus C. D., Manaloto C. O. Dengue fever/dengue haemorrhagic fever in Filipino children: clinical experience during the 1983-1984 epidemic. Southeast Asian J Trop Med Public Health. 1987 Sep;18(3):284–290. [PubMed] [Google Scholar]
  20. Weddle J. R., Chan T. C., Thompson K., Paxton H., Kelly D. J., Dasch G., Strickman D. Effectiveness of a dot-blot immunoassay of anti-Rickettsia tsutsugamushi antibodies for serologic analysis of scrub typhus. Am J Trop Med Hyg. 1995 Jul;53(1):43–46. [PubMed] [Google Scholar]
  21. Weiblen B. J., Schumacher R. T., Hoff R. Detection of IgM and IgA HIV antibodies after removal of IgG with recombinant protein G. J Immunol Methods. 1990 Feb 9;126(2):199–204. doi: 10.1016/0022-1759(90)90151-k. [DOI] [PubMed] [Google Scholar]

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