This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by McSharry, J. M.
Right arrow Articles by Crumpacker, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by McSharry, J. M.
Right arrow Articles by Crumpacker, C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, April 1998, p. 958-964, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Flow Cytometric Determination of Ganciclovir Susceptibilities of Human Cytomegalovirus Clinical Isolates

James M. McSharry,1,* Nell S. Lurain,2 George L. Drusano,1 Alan Landay,2 Jody Manischewitz,3 Mostafa Nokta,4 Maurice O'Gorman,5 Howard M. Shapiro,6 Adriana Weinberg,7 Patricia Reichelderfer,8 and Clyde Crumpacker9

Albany Medical College, Albany, New York 122081; Rush Presbyterian St. Luke's Medical Center, Chicago, Illinois 60153-38332; Food and Drug Administration, Bethesda, Maryland 208923; University of Texas Medical Branch, Galveston, Texas 77555-08354; Northwestern University Children's Memorial Hospital, Chicago, Illinois 606145; Newton, Massachusetts 021656; University of Colorado Medical Center, Denver, Colorado 802627; National Institutes of Health, Bethesda, Maryland 20892-76208; and Beth Israel-Deaconess Medical Center, Boston, Massachusetts 02215-54009

Received 27 August 1997/Returned for modification 17 November 1997/Accepted 5 January 1998

A flow cytometric assay has been developed for the measurement of susceptibilities to ganciclovir of laboratory strains and clinical isolates of human cytomegalovirus (HCMV). The assay uses fluorochrome-labeled monoclonal antibodies to HCMV immediate-early and late antigens to identify HCMV-infected cells and flow cytometry to detect and quantitate the number of antigen-positive cells. By this assay, the 50 and 90% inhibitory concentrations (IC50 and IC90, respectively) of ganciclovir for the AD169 strain of HCMV were 1.7 and 9.2 µM, respectively, and the IC50 for the ganciclovir-resistant D6/3/1 derivative of the AD169 strain was greater than 12 µM. The ganciclovir susceptibilities of 17 HCMV clinical isolates were also determined by flow cytometric analysis of the effect of ganciclovir on late-antigen synthesis in HCMV-infected cells. The average IC50 of ganciclovir for drug-sensitive HCMV clinical isolates was 3.79 µM (±2.60). The plaque-reduction assay for these clinical isolates yielded an average IC50 of 2.80 µM (±1.46). Comparison of the results of the flow cytometry assays with those obtained from the plaque-reduction assays demonstrated acceptable bias and precision. Flow cytometric and plaque-reduction analysis of cells infected with ganciclovir-resistant clinical isolates failed to show a reduction in the percentage of late-antigen-positive cells or PFU, even at 96 µM ganciclovir. The flow cytometric assay for determining ganciclovir susceptibility of HCMV is quantitative, and objective, and potentially automatable, and its results are reproducible among laboratories.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Molecular Genetics, A-68, Albany Medical College, 47 New Scotland Ave., Albany, NY 12208. Phone: (518) 262-5174. Fax: (518) 262-5748. E-mail: jim_mcsharry{at}ccgateway.amc.edu.

Journal of Clinical Microbiology, April 1998, p. 958-964, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Lee, G.-C., Lee, D.-G., Choi, S.-M., Yoo, J.-H., Park, S.-H., Choi, J.-H., Min, W.-S., Cho, O.-H., Lee, C.-H., Shin, W.-S. (2005). Use of Time-Saving Flow Cytometry for Rapid Determination of Resistance of Human Cytomegalovirus to Ganciclovir. J. Clin. Microbiol. 43: 5003-5008 [Abstract] [Full Text]  
  • Gilbert, C., Boivin, G. (2005). Human Cytomegalovirus Resistance to Antiviral Drugs. Antimicrob. Agents Chemother. 49: 873-883 [Full Text]  
  • Chutkowski, C., Olson, B., McDonough, A., Mahoney, J., McSharry, J. J. (2002). Use of a Single Monoclonal Antibody To Determine the Susceptibilities of Herpes Simplex Virus Type 1 and Type 2 Clinical Isolates to Acyclovir. CVI 9: 1379-1381 [Abstract] [Full Text]  
  • McSharry, J. J., McDonough, A., Olson, B., Talarico, C., Davis, M., Biron, K. K. (2001). Inhibition of Ganciclovir-Susceptible and -Resistant Human Cytomegalovirus Clinical Isolates by the Benzimidazole L-Riboside 1263W94. CVI 8: 1279-1281 [Abstract] [Full Text]  
  • Lurain, N. S., Weinberg, A., Crumpacker, C. S., Chou, S. (2001). Sequencing of Cytomegalovirus UL97 Gene for Genotypic Antiviral Resistance Testing. Antimicrob. Agents Chemother. 45: 2775-2780 [Abstract] [Full Text]  
  • McSharry, J. J., McDonough, A., Olson, B., Hallenberger, S., Reefschlaeger, J., Bender, W., Drusano, G. L. (2001). Susceptibilities of Human Cytomegalovirus Clinical Isolates to BAY38-4766, BAY43-9695, and Ganciclovir. Antimicrob. Agents Chemother. 45: 2925-2927 [Abstract] [Full Text]  
  • Alvarez-Barrientos, A., Arroyo, J., Canton, R., Nombela, C., Sanchez-Perez, M. (2000). Applications of Flow Cytometry to Clinical Microbiology. Clin. Microbiol. Rev. 13: 167-195 [Abstract] [Full Text]  
  • Sia, I. G., Patel, R. (2000). New Strategies for Prevention and Therapy of Cytomegalovirus Infection and Disease in Solid-Organ Transplant Recipients. Clin. Microbiol. Rev. 13: 83-121 [Abstract] [Full Text]  
  • Erice, A. (1999). Resistance of Human Cytomegalovirus to Antiviral Drugs. Clin. Microbiol. Rev. 12: 286-297 [Abstract] [Full Text]  
  • McSharry, J. J., Lurain, N. S., Drusano, G. L., Landay, A. L., Notka, M., O'Gorman, M. R. G., Weinberg, A., Shapiro, H. M., Reichelderfer, P. S., Crumpacker, C. S. (1998). Rapid Ganciclovir Susceptibility Assay Using Flow Cytometry for Human Cytomegalovirus Clinical Isolates. Antimicrob. Agents Chemother. 42: 2326-2331 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»