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. 1995 Jun;33(6):1646–1649. doi: 10.1128/jcm.33.6.1646-1649.1995

Comparison of several fixation methods for cytomegalovirus antigenemia assay.

J L Pérez 1, M De Oña 1, J Niubò 1, H Villar 1, S Melón 1, A García 1, R Martín 1
PMCID: PMC228235  PMID: 7650205

Abstract

Two fixation methods based on formaldehyde or acetone for qualitative cytomegalovirus antigenemia assay were evaluated on 405 consecutive blood samples. Cytomegalovirus was detected in 40 samples by the antigenemia assay: 36 were detected by formaldehyde fixation; 22, by acetone; and 18, by both methods. Differences were statistically significant (P = 0.0043). In addition, four fixation methods (two based on formalin [with and without permeabilization] and two using acetone at different fixation times) for quantitative antigenemia assay in a different set of 32 samples from known viremic patients were evaluated. Formalin-based methods were superior to acetone-based methods, showing statistically significant differences in either the number of positive samples detected (P < 0.02; McNemar test) or the mean positive cell counts (P < 0.003; two-tailed Student's t test for paired samples). No differences between the two formalin-based methods were found. We recommend the formaldehyde fixation procedure without subsequent permeabilization because of its simplicity and sensitivity.

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Selected References

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  1. Aguado S., Gómez E., Rodríguez A., Martínez A., Oña M., Alvarez-Grande J. A prospective study of a rapid method for diagnosing cytomegalovirus infections in immunosuppressed patients. Nephron. 1989;53(4):322–324. doi: 10.1159/000185775. [DOI] [PubMed] [Google Scholar]
  2. Bein G., Bitsch A., Hoyer J., Kirchner H. The detection of human cytomegalovirus immediate early antigen in peripheral blood leucocytes. J Immunol Methods. 1991 Mar 21;137(2):175–180. doi: 10.1016/0022-1759(91)90022-8. [DOI] [PubMed] [Google Scholar]
  3. Boeckh M., Woogerd P. M., Stevens-Ayers T., Ray C. G., Bowden R. A. Factors influencing detection of quantitative cytomegalovirus antigenemia. J Clin Microbiol. 1994 Mar;32(3):832–834. doi: 10.1128/jcm.32.3.832-834.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dankner W. M., McCutchan J. A., Richman D. D., Hirata K., Spector S. A. Localization of human cytomegalovirus in peripheral blood leukocytes by in situ hybridization. J Infect Dis. 1990 Jan;161(1):31–36. doi: 10.1093/infdis/161.1.31. [DOI] [PubMed] [Google Scholar]
  5. Erice A., Holm M. A., Gill P. C., Henry S., Dirksen C. L., Dunn D. L., Hillam R. P., Balfour H. H., Jr Cytomegalovirus (CMV) antigenemia assay is more sensitive than shell vial cultures for rapid detection of CMV in polymorphonuclear blood leukocytes. J Clin Microbiol. 1992 Nov;30(11):2822–2825. doi: 10.1128/jcm.30.11.2822-2825.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Freymuth F., Gennetay E., Petitjean J., Eugene G., Hurault de Ligny B., Ryckelynck J. P., Legoff C., Hazera P., Bazin C. Comparison of nested PCR for detection of DNA in plasma with pp65 leukocytic antigenemia procedure for diagnosis of human cytomegalovirus infection. J Clin Microbiol. 1994 Jun;32(6):1614–1618. doi: 10.1128/jcm.32.6.1614-1618.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gerna G., Revello M. G., Percivalle E., Morini F. Comparison of different immunostaining techniques and monoclonal antibodies to the lower matrix phosphoprotein (pp65) for optimal quantitation of human cytomegalovirus antigenemia. J Clin Microbiol. 1992 May;30(5):1232–1237. doi: 10.1128/jcm.30.5.1232-1237.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gleaves C. A., Smith T. F., Shuster E. A., Pearson G. R. Comparison of standard tube and shell vial cell culture techniques for the detection of cytomegalovirus in clinical specimens. J Clin Microbiol. 1985 Feb;21(2):217–221. doi: 10.1128/jcm.21.2.217-221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gómez E., Aguado S., Melón S., Martínez A., de Oña M., Cimadevilla R., Gorostidi M., González E., Alvarez Grande J. A prospective study of the usefulness of rapid diagnostic tests in monitoring cytomegalovirus infection in renal transplants. Transplant Proc. 1992 Dec;24(6):2631–2633. [PubMed] [Google Scholar]
  10. Koskinen P. K., Nieminen M. S., Mattila S. P., Häyry P. J., Lautenschlager I. T. The correlation between symptomatic CMV infection and CMV antigenemia in heart allograft recipients. Transplantation. 1993 Mar;55(3):547–551. doi: 10.1097/00007890-199303000-00017. [DOI] [PubMed] [Google Scholar]
  11. Landry M. L., Ferguson D. Comparison of quantitative cytomegalovirus antigenemia assay with culture methods and correlation with clinical disease. J Clin Microbiol. 1993 Nov;31(11):2851–2856. doi: 10.1128/jcm.31.11.2851-2856.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meyers J. D., Ljungman P., Fisher L. D. Cytomegalovirus excretion as a predictor of cytomegalovirus disease after marrow transplantation: importance of cytomegalovirus viremia. J Infect Dis. 1990 Aug;162(2):373–380. doi: 10.1093/infdis/162.2.373. [DOI] [PubMed] [Google Scholar]
  13. Niubò J., Pérez J. L., Carvajal A., Ardanuy C., Martín R. Effect of delayed processing of blood samples on performance of cytomegalovirus antigenemia assay. J Clin Microbiol. 1994 Apr;32(4):1119–1120. doi: 10.1128/jcm.32.4.1119-1120.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pérez J. L., Mariscal D., Tubau F., Niubò J., Martín R. Evaluation of the CMV-vue antigenemia assay for rapid detection of cytomegalovirus in peripheral blood leukocytes. Diagn Microbiol Infect Dis. 1994 May;19(1):15–18. doi: 10.1016/0732-8893(94)90045-0. [DOI] [PubMed] [Google Scholar]
  15. Revello M. G., Percivalle E., Zavattoni M., Parea M., Grossi P., Gerna G. Detection of human cytomegalovirus immediate early antigen in leukocytes as a marker of viremia in immunocompromised patients. J Med Virol. 1989 Oct;29(2):88–93. doi: 10.1002/jmv.1890290204. [DOI] [PubMed] [Google Scholar]
  16. Storch G. A., Buller R. S., Bailey T. C., Ettinger N. A., Langlois T., Gaudreault-Keener M., Welby P. L. Comparison of PCR and pp65 antigenemia assay with quantitative shell vial culture for detection of cytomegalovirus in blood leukocytes from solid-organ transplant recipients. J Clin Microbiol. 1994 Apr;32(4):997–1003. doi: 10.1128/jcm.32.4.997-1003.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. The T. H., van der Ploeg M., van den Berg A. P., Vlieger A. M., van der Giessen M., van Son W. J. Direct detection of cytomegalovirus in peripheral blood leukocytes--a review of the antigenemia assay and polymerase chain reaction. Transplantation. 1992 Aug;54(2):193–198. doi: 10.1097/00007890-199208000-00001. [DOI] [PubMed] [Google Scholar]
  18. van den Berg A. P., van der Bij W., van Son W. J., Anema J., van der Giessen M., Schirm J., Tegzess A. M., The T. H. Cytomegalovirus antigenemia as a useful marker of symptomatic cytomegalovirus infection after renal transplantation--a report of 130 consecutive patients. Transplantation. 1989 Dec;48(6):991–995. doi: 10.1097/00007890-198912000-00019. [DOI] [PubMed] [Google Scholar]
  19. van der Bij W., van Dijk R. B., van Son W. J., Torensma R., Prenger K. B., Prop J., Tegzess A. M., The T. H. Antigen test for early diagnosis of active cytomegalovirus infection in heart transplant recipients. J Heart Transplant. 1988 Mar-Apr;7(2):106–109. [PubMed] [Google Scholar]
  20. van der Giessen M., The T. H., van Son W. J. Cytomegalovirus antigenemia assay. J Clin Microbiol. 1991 Dec;29(12):2909–2910. doi: 10.1128/jcm.29.12.2909-2910.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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