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Antimicrobial Agents and Chemotherapy, September 1998, p. 2326-2331, Vol. 42, No. 9
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Rapid Ganciclovir Susceptibility Assay Using Flow Cytometry for Human Cytomegalovirus Clinical Isolates

James J. McSharry,1,* Nell S. Lurain,2 George L. Drusano,1 Alan L. Landay,2 Mostafa Notka,3 Maurice R. G. O'Gorman,4 Adriana Weinberg,5 Howard M. Shapiro,6 Patricia S. Reichelderfer,7 and Clyde S. Crumpacker8

Albany Medical College, Albany, New York 122081; Rush Presbyterian St. Luke's Medical Center, Chicago, Illinois 60612-38332; University of Texas Medical Branch, Galveston, Texas 77555-08353; Northwestern University Children's Memorial Hospital, Chicago, Illinois 606144; University of Colorado Medical Center, Denver, Colorado 802625; Newton, Massachusetts 026156; National Institutes of Health, Bethesda, Maryland 20892-76207; and Beth Israel-Deaconess Medical Center, Boston, Massachusetts 02215-54008

Received 23 February 1998/Returned for modification 16 April 1998/Accepted 12 May 1998

Rapid, quantitative, and objective determination of the susceptibilities of human cytomegalovirus (HCMV) clinical isolates to ganciclovir has been assessed by an assay that uses a fluorochrome-labeled monoclonal antibody to an HCMV immediate-early antigen and flow cytometry. Analysis of the ganciclovir susceptibilities of 25 phenotypically characterized clinical isolates by flow cytometry demonstrated that the 50% inhibitory concentrations (IC50s) of ganciclovir for 19 of the isolates were between 1.14 and 6.66 µM, with a mean of 4.32 µM (±1.93) (sensitive; IC50 less than 7 µM), the IC50s for 2 isolates were 8.48 and 9.79 µM (partially resistant), and the IC50s for 4 isolates were greater than 96 µM (resistant). Comparative analysis of the drug susceptibilities of these clinical isolates by the plaque reduction assay gave IC50s of less than 6 µM, with a mean of 2.88 µM (±1.40) for the 19 drug-sensitive isolates, IC50s of 6 to 8 µM for the partially resistant isolates, and IC50s of greater than 12 µM for the four resistant clinical isolates. Comparison of the IC50s for the drug-susceptible and partially resistant clinical isolates obtained by the flow cytometry assay with the IC50s obtained by the plaque reduction assay showed an acceptable correlation (r2 = 0.473; P = 0.001), suggesting that the flow cytometry assay could substitute for the more labor-intensive, subjective, and time-consuming plaque reduction assay.

* 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.

Antimicrobial Agents and Chemotherapy, September 1998, p. 2326-2331, Vol. 42, No. 9
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.