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J Clin Virol. 2007 Feb;38(2):139-45. Epub 2006 Dec 13.

Factors that affect in vitro measurement of the susceptibility of herpes simplex virus to nucleoside analogues.

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Pediatric Infectious Diseases, University of Colorado Health Sciences Center, Denver, CO, United States.



To identify factors that contribute to variability of HSV antiviral susceptibility breakpoints.


Acyclovir and penciclovir IC(50)'s for 12 HSV clinical isolates were measured in two laboratories using plaque reduction assay (PRA), an enzyme immunoassay (EIA)-based antigen reduction, and DNA hybridization on Vero, A549, MRC-5, HEL299 and HELG monolayers. Pair-wise comparisons were performed to evaluate variables including testing laboratory, technique, monolayer, and antiviral. The proportion of false results was analyzed using a conventional susceptibility IC(50) breakpoint of 2 microg/ml.


Acyclovir-resistant HSV isolates were correctly identified by all methods. In contrast, there were 6-67% of susceptible isolates incorrectly characterized as drug-resistant. Variables associated with these errors included testing site, assay method, cell line and antiviral. A549, DNA hybridization, and penciclovir were associated with the highest IC(50)'s, whereas the PRA, EIA, and human fibroblast-monolayers provided the best differentiation between susceptible and resistant HSV isolates.


The current recommendations to use a single discriminating value to define HSV resistance to nucleoside analogues can be problematic. False results are influenced in various degrees by the laboratory method, tissue culture and antivirals.

[Indexed for MEDLINE]

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