Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 Aug;27(8):1810–1813. doi: 10.1128/jcm.27.8.1810-1813.1989

Immunofluorescence assay for human immunodeficiency virus antibody: investigation of cell fixation for virus inactivation and antigen preservation.

M Fauvel 1, G Ozanne 1
PMCID: PMC267676  PMID: 2671031

Abstract

Four cell fixation procedures were investigated for their abilities to inactivate human immunodeficiency virus (HIV) and preserve its antigenicity for antibody detection by immunofluorescence in MOLT-4-T4 cells. Air-dried cell smears were fixed in cold acetone, in acetone-methanol (1:1), in acetone-methanol (1:1) followed by 70% ethanol and then methanol, or in paraformaldehyde-acetone. Acetone alone did not inactivate cell-associated HIV, but the other three procedures did. HIV inactivation was achieved by storage of acetone-fixed cells at -70 degrees for 40 days. Antigenicity was measured by immunofluorescence assay titrations of selected human sera, a cerebrospinal fluid, and a gp41 monoclonal antibody. Acetone provided the best fixation as measured by fluorescence intensity and antibody titers. The other fixation methods all yielded weaker fluorescence signals and/or decreased titers. Acetone fixation and storage for 40 days at -70 degrees C provides safe and accurate immunofluorescence assay reagents.

Full text

PDF
1810

Images in this article

1812Image
on p.1812

Selected References

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

  1. Barin F., McLane M. F., Allan J. S., Lee T. H., Groopman J. E., Essex M. Virus envelope protein of HTLV-III represents major target antigen for antibodies in AIDS patients. Science. 1985 May 31;228(4703):1094–1096. doi: 10.1126/science.2986291. [DOI] [PubMed] [Google Scholar]
  2. Blumberg R. S., Sandstrom E. G., Paradis T. J., Neumeyer D. N., Sarngadharan M. G., Hartshorn K. L., Byington R. E., Hirsch M. S., Schooley R. T. Detection of human T-cell lymphotropic virus type III-related antigens and anti-human T-cell lymphotropic virus type III antibodies by anticomplementary immunofluorescence. J Clin Microbiol. 1986 Jun;23(6):1072–1077. doi: 10.1128/jcm.23.6.1072-1077.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carlson J. R., Yee J., Hinrichs S. H., Bryant M. L., Gardner M. B., Pedersen N. C. Comparison of indirect immunofluorescence and Western blot for detection of anti-human immunodeficiency virus antibodies. J Clin Microbiol. 1987 Mar;25(3):494–497. doi: 10.1128/jcm.25.3.494-497.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gaines H., von Sydow M., Sönnerborg A., Albert J., Czajkowski J., Pehrson P. O., Chiodi F., Moberg L., Fenyö E. M., Asjö B. Antibody response in primary human immunodeficiency virus infection. Lancet. 1987 May 30;1(8544):1249–1253. doi: 10.1016/s0140-6736(87)92696-1. [DOI] [PubMed] [Google Scholar]
  5. Gallo D., Diggs J. L., Shell G. R., Dailey P. J., Hoffman M. N., Riggs J. L. Comparison of detection of antibody to the acquired immune deficiency syndrome virus by enzyme immunoassay, immunofluorescence, and Western blot methods. J Clin Microbiol. 1986 Jun;23(6):1049–1051. doi: 10.1128/jcm.23.6.1049-1051.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hedenskog M., Dewhurst S., Ludvigsen C., Sinangil F., Rodriguez L., Wu Y. T., Volsky D. J. Testing for antibodies to AIDS-associated retrovirus (HTLV-III/LAV) by indirect fixed cell immunofluorescence: specificity, sensitivity, and applications. J Med Virol. 1986 Aug;19(4):325–334. doi: 10.1002/jmv.1890190405. [DOI] [PubMed] [Google Scholar]
  7. Hoffman A. D., Banapour B., Levy J. A. Characterization of the AIDS-associated retrovirus reverse transcriptase and optimal conditions for its detection in virions. Virology. 1985 Dec;147(2):326–335. doi: 10.1016/0042-6822(85)90135-7. [DOI] [PubMed] [Google Scholar]
  8. Imrie A. A., Kehrer S., Smith G. W., Penny R., Cooper D. A. Seroimmunology of AIDS retrovirus infection. I. Use of immunofluorescence assay to confirm sera with ELISA reactivity. Pathology. 1986 Oct;18(4):438–443. doi: 10.3109/00313028609087565. [DOI] [PubMed] [Google Scholar]
  9. Jackson J. B., Balfour H. H., Jr Practical diagnostic testing for human immunodeficiency virus. Clin Microbiol Rev. 1988 Jan;1(1):124–138. doi: 10.1128/cmr.1.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lennette E. T., Karpatkin S., Levy J. A. Indirect immunofluorescence assay for antibodies to human immunodeficiency virus. J Clin Microbiol. 1987 Feb;25(2):199–202. doi: 10.1128/jcm.25.2.199-202.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lifson J. D., Sasaki D. T., Engleman E. G. Utility of formaldehyde fixation for flow cytometry and inactivation of the AIDS associated retrovirus. J Immunol Methods. 1986 Jan 22;86(1):143–149. doi: 10.1016/0022-1759(86)90278-4. [DOI] [PubMed] [Google Scholar]
  12. McHugh T. M., Stites D. P., Casavant C. H., Carlson J. R., Yee J., McVay P. A., Busch M. P., Levy J. A. Evaluation of the indirect immunofluorescence assay as a confirmatory test for detecting antibodies to the human immunodeficiency virus. Diagn Immunol. 1986;4(5):233–240. [PubMed] [Google Scholar]
  13. Ozanne G., Fauvel M. Performance and reliability of five commercial enzyme-linked immunosorbent assay kits in screening for anti-human immunodeficiency virus antibody in high-risk subjects. J Clin Microbiol. 1988 Aug;26(8):1496–1500. doi: 10.1128/jcm.26.8.1496-1500.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pan L. Z., Cheng-Mayer C., Levy J. A. Patterns of antibody response in individuals infected with the human immunodeficiency virus. J Infect Dis. 1987 Apr;155(4):626–632. doi: 10.1093/infdis/155.4.626. [DOI] [PubMed] [Google Scholar]
  15. Resnick L., Veren K., Salahuddin S. Z., Tondreau S., Markham P. D. Stability and inactivation of HTLV-III/LAV under clinical and laboratory environments. JAMA. 1986 Apr 11;255(14):1887–1891. [PubMed] [Google Scholar]
  16. Saah A. J., Farzadegan H., Fox R., Nishanian P., Rinaldo C. R., Jr, Phair J. P., Fahey J. L., Lee T. H., Polk B. F. Detection of early antibodies in human immunodeficiency virus infection by enzyme-linked immunosorbent assay, Western blot, and radioimmunoprecipitation. J Clin Microbiol. 1987 Sep;25(9):1605–1610. doi: 10.1128/jcm.25.9.1605-1610.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sandstrom E. G., Schooley R. T., Ho D. D., Byington R., Sarngadharan M. G., MacLane M. E., Essex M., Gallo R. C., Hirsch M. S. Detection of human anti-HTLV-III antibodies by indirect immunofluorescence using fixed cells. Transfusion. 1985 Jul-Aug;25(4):308–312. doi: 10.1046/j.1537-2995.1985.25485273806.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES