Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1989 Jun;27(6):1159–1162. doi: 10.1128/jcm.27.6.1159-1162.1989

Clinical experience with cytomegalovirus isolation using conventional cell cultures and early antigen detection in centrifugation-enhanced shell vial cultures.

D S Leland 1, R L Hansing 1, M L French 1
PMCID: PMC267519  PMID: 2546969

Abstract

A total of 1,915 clinical samples was inoculated by low-speed centrifugation into shell vials (Bartels Immunodiagnostics, Bellvue, Wash.) containing cover slip monolayers of MRC-5 fibroblasts. At 1 and 2 days postinoculation, one cover slip was stained by an indirect immunofluorescence technique using a monoclonal antibody (Biotech Research Laboratories for Dupont, Billerica, Mass.) to cytomegalovirus (CMV) early antigen (EA). Clinical samples were also inoculated into three MRC-5 or MRHF cell cultures which were observed for 30 days for the appearance of a cytopathic effect (CPE). Of 157 CMV-positive samples, 92 (59%) were identified by centrifugation-enhanced EA (CE-EA) and 131 (83%) produced a CPE. CE-EA was less sensitive than CPE for all types of samples, although 17% of CMV-positive samples were detected by CE-EA alone. Evaluation of the CMV status of patients with CE-EA-positive-CPE-negative samples indicated that these samples likely represented true CMV-positive results. The average elapsed time between culture inoculation and identification of CMV decreased as follows when both CE-EA and CPE, rather than CPE alone, were used: urines, 15 to 7 days; buffy coats, 18 to 9 days; lung samples, 13 to 8 days; throat samples, 18 to 7 days. Although CE-EA was less sensitive than 30-day cell culture, both CE-EA and CPE were identified as valuable in CMV detection, and neither could be discontinued without a decrease in the CMV isolation rate or an increase in the turnaround time.

Full text

PDF
1159

Selected References

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

  1. Alpert G., Mazeron M. C., Colimon R., Plotkin S. Rapid detection of human cytomegalovirus in the urine of humans. J Infect Dis. 1985 Sep;152(3):631–633. doi: 10.1093/infdis/152.3.631. [DOI] [PubMed] [Google Scholar]
  2. DeGirolami P. C., Dakos J., Eichelberger K., Mills L. S., DeLuca A. M. Rapid detection of cytomegalovirus in clinical specimens by immunofluorescent staining of shell vial cultures. Am J Clin Pathol. 1988 Apr;89(4):528–532. doi: 10.1093/ajcp/89.4.528. [DOI] [PubMed] [Google Scholar]
  3. Gleaves C. A., Lee C. F., Kirsch L., Meyers J. D. Evaluation of a direct fluorescein-conjugated monoclonal antibody for detection of cytomegalovirus in centrifugation culture. J Clin Microbiol. 1987 Aug;25(8):1548–1550. doi: 10.1128/jcm.25.8.1548-1550.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Gleaves C. A., Smith T. F., Shuster E. A., Pearson G. R. Rapid detection of cytomegalovirus in MRC-5 cells inoculated with urine specimens by using low-speed centrifugation and monoclonal antibody to an early antigen. J Clin Microbiol. 1984 Jun;19(6):917–919. doi: 10.1128/jcm.19.6.917-919.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Paya C. V., Wold A. D., Ilstrup D. M., Smith T. F. Evaluation of number of shell vial cell cultures per clinical specimen for rapid diagnosis of cytomegalovirus infection. J Clin Microbiol. 1988 Feb;26(2):198–200. doi: 10.1128/jcm.26.2.198-200.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Shuster E. A., Beneke J. S., Tegtmeier G. E., Pearson G. R., Gleaves C. A., Wold A. D., Smith T. F. Monoclonal antibody for rapid laboratory detection of cytomegalovirus infections: characterization and diagnostic application. Mayo Clin Proc. 1985 Sep;60(9):577–585. doi: 10.1016/s0025-6196(12)60979-3. [DOI] [PubMed] [Google Scholar]
  9. Stirk P. R., Griffiths P. D. Use of monoclonal antibodies for the diagnosis of cytomegalovirus infection by the detection of early antigen fluorescent foci (DEAFF) in cell culture. J Med Virol. 1987 Apr;21(4):329–337. doi: 10.1002/jmv.1890210405. [DOI] [PubMed] [Google Scholar]

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

RESOURCES