Journal
JOURNAL OF CLINICAL VIROLOGY
Volume 56, Issue 1, Pages 19-24Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jcv.2012.09.004
Keywords
Herpes simplex virus type 1; Aciclovir; Foscarnet; Susceptibility testing; Real time PCR
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Background: Susceptibility testing of herpes simplex virus type 1 (HSV-1) is traditionally performed by a plaque reduction assay (PRA), but this is labor intensive, time consuming and has a manual read out. Objectives: The goal of this study was to develop an internally controlled real time PCR-based phenotypical susceptibility test for HSV-1 that is suitable for use in a clinical diagnostic setting. Study design: A DNA reduction assay (DRA) was developed and validated on a test panel of 26 well-characterized isolates of varying susceptibility to aciclovir or foscarnet, including low-level resistant isolates. The DRA consisted of pre-culture of a clinical sample for 48 h and subsequent culture in the presence of antivirals for 24 h. Viral DNA concentration in the culture lysates was measured by an internally controlled quantitative real-time HSV-1 PCR and corrected for cell count and lysis by beta-globin PCR. DRA results were compared to results from PRA and sequence analysis. Results: DRA results were in accordance with PRA results for both aciclovir and foscarnet susceptibility and appeared to have good discriminative value for low-level resistance due to UL30 gene mutations. Although the direct application of DRA in clinical samples appeared not possible, short pre-culture of 48 h was sufficient and ensured results within a clinically relevant time frame of 5 days. Conclusions: DRA is an accurate, rapid and easy to perform phenotypical susceptibility test for HSV-1. (C) 2012 Elsevier B.V. All rights reserved.
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