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Journal of Clinical Microbiology, October 2005, p. 5003-5008, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5003-5008.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of Time-Saving Flow Cytometry for Rapid Determination of Resistance of Human Cytomegalovirus to Ganciclovir

Gyu-Cheol Lee,1,2 Dong-Gun Lee,1 Su-Mi Choi,1 Jin-Hong Yoo,1 Sun-Hee Park,1 Jung-Hyun Choi,1 Woo-Sung Min,1 Ok-Hee Cho,1 Chan-Hee Lee,3 and Wan-Shik Shin1*

The Clinical Research Institute, St. Mary's Hospital, The Catholic Hemopoietic Stem Cell Transplantation Center, The Catholic University of Korea, College of Medicine, Seoul 150-713, Korea,1 Water Quality Analysis Team I, International Water Analysis Center, Korea Water Resources Corporation, Daejeon 306-711, Korea,2 Department of Microbiology, Natural Sciences College, Chungbuk National University, Cheongju 361-763, Korea3

Received 6 December 2004/ Returned for modification 28 January 2005/ Accepted 1 August 2005

There are two ways to assess the susceptibility of human cytomegalovirus (HCMV) to ganciclovir (GCV): one is a genotypic test that detects resistance-related mutations and the other is a phenotypic test that actually assesses susceptibility. The advantages of genotyping the UL97 gene are its rapidity and accuracy. However, to detect novel mutations or mutations affecting the UL54 DNA polymerase, a phenotypic test such as the plaque reduction assay (PRA) is also required. To avoid the shortcomings of PRA such as its time-consuming nature and labor-intensiveness, we developed a time-saving fluorescence-activated cell sorting (TS-FACS) technique. We obtained a GCV 50% inhibitory concentration (IC50) from five clinical isolates and an HCMV laboratory strain (AD169) and compared the results with those from the PRA. The laboratory strain and three clinical isolates were sensitive to GCV. Although there was a minor discrepancy in the case of one of the three isolates, the GCV IC50 values obtained by TS-FACS analysis correlated well with the results of the PRA. The remaining two isolates were resistant to GCV; one was GCV resistant due to the mutation M460V, and the GCV IC50 results obtained by TS-FACS analysis and by PRA were also comparable. The advantages of TS-FACS analysis are the shorter time required, the possibility of automation, and its comparability to PRA, considered the gold standard. Thus, TS-FACS analysis may be useful as an alternative to PRA in the clinic.

* Corresponding author. Mailing address: St. Mary's Hospital, #62, Yoido-dong, Yongdungpo-gu, 150-713, Seoul, Korea. Phone: 82-2-3779-1338. Fax: 82-2-780-9114. E-mail: devilsoldier{at}hanmail.net.

Journal of Clinical Microbiology, October 2005, p. 5003-5008, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5003-5008.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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