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. 1989 Jul;27(7):1623–1625. doi: 10.1128/jcm.27.7.1623-1625.1989

Double-label immunofluorescence method for simultaneous detection of adenovirus and herpes simplex virus from the eye.

P Walpita 1, S Darougar 1
PMCID: PMC267626  PMID: 2549090

Abstract

The development and application of a double-label immunofluorescence method which has the potential to screen for single or dual infections from any site, in single shell vial cultures, is described. In this study, a total of 1,141 ocular specimens were inoculated in shell vials, centrifuged at 15,000 X g for 1 h, incubated at 37 degrees C for 48 h, and fixed in methanol at room temperature for 15 min. The virus inclusions were detected by staining with a double-label indirect immunofluorescence procedure using mixtures of appropriate first antibodies, followed by fluorescein- and rhodamine-conjugated second antibodies. Each specimen was also inoculated in parallel by the conventional virus isolation method. The sensitivity and specificity of the double-label shell vial procedure were comparable to those with the conventional method, and the former test took only 48 h to complete. The test offers a rapid and simple single-vial procedure which allows for individual or simultaneous detection of multiple pathogens. It results in savings in time and cost over the conventional virus isolation method and other shell vial procedures.

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Selected References

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

  1. Ayliffe M. J. Double staining immunofluorescence procedure for enumeration of cells containing cytoplasmic immunoglobulin in human bone marrow: 40 selected cases. J Clin Pathol. 1985 Sep;38(9):989–994. doi: 10.1136/jcp.38.9.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bologna M., Allen R., Dulbecco R. A method for double immunofluorescent staining by the indirect procedure with antibodies of the same isotype. J Immunol Methods. 1986 Jan 22;86(1):151–153. doi: 10.1016/0022-1759(86)90279-6. [DOI] [PubMed] [Google Scholar]
  3. Darougar S., Gibson J. A., Thaker U. Effect of centrifugation on herpes simplex virus isolation. J Med Virol. 1981;8(4):231–235. doi: 10.1002/jmv.1890080403. [DOI] [PubMed] [Google Scholar]
  4. Darougar S., Walpita P., Thaker U., Viswalingam N., Gardner L., McSwiggan D. A. Adenovirus serotypes isolated from ocular infections in London. Br J Ophthalmol. 1983 Feb;67(2):111–114. doi: 10.1136/bjo.67.2.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Evans R. T., Woodland R. M. Detection of chlamydiae by isolation and direct examination. Br Med Bull. 1983 Apr;39(2):181–186. doi: 10.1093/oxfordjournals.bmb.a071813. [DOI] [PubMed] [Google Scholar]
  6. Gordon F. B., Harper I. A., Quan A. L., Treharne J. D., Dwyer R. S., Garland J. A. Detection of Chlamydia (Bedsonia) in certain infections of man. I. Laboratory procedures: comparison of yolk sac and cell culture for detection and isolation. J Infect Dis. 1969 Oct;120(4):451–462. doi: 10.1093/infdis/120.4.451. [DOI] [PubMed] [Google Scholar]
  7. Johnson G. D., Davidson R. S., McNamee K. C., Russell G., Goodwin D., Holborow E. J. Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy. J Immunol Methods. 1982 Dec 17;55(2):231–242. doi: 10.1016/0022-1759(82)90035-7. [DOI] [PubMed] [Google Scholar]
  8. Liu C., Llanes-Rodas R. Application of the immunofluorescent technic to the study of pathogenesis and rapid diagnosis of viral infections. Am J Clin Pathol. 1972 Jun;57(6):829–834. doi: 10.1093/ajcp/57.6.829. [DOI] [PubMed] [Google Scholar]
  9. Ozturk G. E., Hayward A. R., Weil R., 3rd, Kohler P. F. Double fluorescence technique for measurement of complement-fixing antibody to lymphocyte subsets. J Immunol Methods. 1985 Dec 17;85(1):163–168. doi: 10.1016/0022-1759(85)90284-4. [DOI] [PubMed] [Google Scholar]
  10. Paya C. V., Wold A. D., Smith T. F. Detection of cytomegalovirus infections in specimens other than urine by the shell vial assay and conventional tube cell cultures. J Clin Microbiol. 1987 May;25(5):755–757. doi: 10.1128/jcm.25.5.755-757.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Schwartz H. S., Vastine D. W., Yamashiroya H., West C. E. Immunofluorescent detection of adenovirus antigen in epidemic keratoconjunctivitis. Invest Ophthalmol. 1976 Mar;15(3):199–207. [PubMed] [Google Scholar]
  12. Walpita P., Darougar S., Thaker U. A rapid and sensitive culture test for detecting herpes simplex virus from the eye. Br J Ophthalmol. 1985 Sep;69(9):637–639. doi: 10.1136/bjo.69.9.637. [DOI] [PMC free article] [PubMed] [Google Scholar]

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