Abstract
Histopaque-1119 (Sigma Chemical Co., St. Louis, Mo.) and Plasmagel (Cellular Products, Inc., Buffalo, N.Y.) were compared as density gradient separation reagents for the separation of polymorphonuclear leukocytes and mononuclear cells from blood from the isolation of cytomegalovirus (CMV). Of 200 peripheral blood specimens examined, CMV was recovered from 51 by both methods. The time of detection of immunofluorescent sites or a cytopathic effect associated with CMV was similar by each method. The Histopaque-1119 method was less time-consuming than the Plasmagel method since it did not require a precentrifugation step for the settling of erythrocytes. The use of Histopaque-1119 will permit an effective alternative single-step method for the separation of blood leukocytes for the isolation of CMV.
Full text
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Dowling J. N., Saslow A. R., Armstrong J. A., Ho M. Cytomegalovirus infection in patients receiving immunosuppressive therapy for rheumatologic disorders. J Infect Dis. 1976 Apr;133(4):399–408. doi: 10.1093/infdis/133.4.399. [DOI] [PubMed] [Google Scholar]
- Einhorn L., Ost A. Cytomegalovirus infection of human blood cells. J Infect Dis. 1984 Feb;149(2):207–214. doi: 10.1093/infdis/149.2.207. [DOI] [PubMed] [Google Scholar]
- English D., Andersen B. R. Single-step separation of red blood cells. Granulocytes and mononuclear leukocytes on discontinuous density gradients of Ficoll-Hypaque. J Immunol Methods. 1974 Aug;5(3):249–252. doi: 10.1016/0022-1759(74)90109-4. [DOI] [PubMed] [Google Scholar]
- Ferrante A., Thong Y. H. Optimal conditions for simultaneous purification of mononuclear and polymorphonuclear leucocytes from human blood by the Hypaque-Ficoll method. J Immunol Methods. 1980;36(2):109–117. doi: 10.1016/0022-1759(80)90036-8. [DOI] [PubMed] [Google Scholar]
- Gadler H., Tillegård A., Groth C. G. Studies of cytomegalovirus infection in renal allograft recipients. Scand J Infect Dis. 1982;14(2):81–87. doi: 10.3109/inf.1982.14.issue-2.01. [DOI] [PubMed] [Google Scholar]
- Howell C. L., Miller M. J., Martin W. J. Comparison of rates of virus isolation from leukocyte populations separated from blood by conventional and Ficoll-Paque/Macrodex methods. J Clin Microbiol. 1979 Oct;10(4):533–537. doi: 10.1128/jcm.10.4.533-537.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mogensen C. E. The glomerular permeability determined by dextran clearance using Sephadex gel filtration. Scand J Clin Lab Invest. 1968;21(1):77–82. doi: 10.3109/00365516809076979. [DOI] [PubMed] [Google Scholar]
- Paya C. V., Wold A. D., Smith T. F. Detection of cytomegalovirus from blood leukocytes separated by sepracell-MN and Ficoll-Paque/Macrodex methods. J Clin Microbiol. 1988 Oct;26(10):2031–2033. doi: 10.1128/jcm.26.10.2031-2033.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rinaldo C. R., Jr, Black P. H., Hirsch M. S. Interaction of cytomegalovirus with leukocytes from patients with mononucleosis due to cytomegalovirus. J Infect Dis. 1977 Nov;136(5):667–678. doi: 10.1093/infdis/136.5.667. [DOI] [PubMed] [Google Scholar]
- Schirm J., Timmerije W., van der Bij W., The T. H., Wilterdink J. B., Tegzess A. M., van Son W. J., Schröder F. P. Rapid detection of infectious cytomegalovirus in blood with the aid of monoclonal antibodies. J Med Virol. 1987 Sep;23(1):31–40. doi: 10.1002/jmv.1890230105. [DOI] [PubMed] [Google Scholar]
- Woods G. L., Proffitt M. R. Comparison of Plasmagel with LeucoPREP-Macrodex methods for separation of leukocytes for virus isolation. Diagn Microbiol Infect Dis. 1987 Oct;8(2):123–126. doi: 10.1016/0732-8893(87)90160-x. [DOI] [PubMed] [Google Scholar]