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Related Experiment Videos

Improved DNA hybridization method for detection of acyclovir-resistant herpes simplex virus.

E M Swierkosz1, D R Scholl, J L Brown

  • 1Department of Pediatrics/Adolescent Medicine, St. Louis University School of Medicine, Missouri.

Antimicrobial Agents and Chemotherapy
|October 1, 1987
PubMed
Summary

A new DNA hybridization assay quickly detects acyclovir-resistant herpes simplex virus (HSV) isolates. This method simplifies antiviral susceptibility testing for clinical laboratories.

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Area of Science:

  • Virology
  • Molecular Biology
  • Clinical Diagnostics

Background:

  • Acyclovir is a primary antiviral medication for herpes simplex virus (HSV) infections.
  • Emergence of acyclovir-resistant HSV isolates necessitates reliable and rapid detection methods.
  • Current susceptibility testing can be time-consuming, impacting timely treatment decisions.

Purpose of the Study:

  • To develop and validate a simplified DNA hybridization method for detecting acyclovir-resistant HSV.
  • To compare the performance of the new DNA hybridization assay with traditional plaque reduction assays.
  • To establish an objective and efficient assay for antiviral susceptibility testing in diagnostic settings.

Main Methods:

  • Herpes simplex virus-infected cell cultures were treated with varying concentrations of acyclovir.

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  • A one-step lysis procedure was used to prepare cell lysates for membrane absorption.
  • Radioiodinated HSV-specific probes were employed for DNA hybridization, followed by washing and gamma counting.
  • Assay performance time was recorded, and results were compared with plaque reduction assays.

    • Main Results:

    • The DNA hybridization assay successfully differentiated acyclovir-resistant (thymidine kinase-negative) from susceptible (thymidine kinase-positive) HSV isolates.
    • Acyclovir-resistant isolates showed a mean 50% inhibitory dose (ID50) greater than 2 µg/ml.
    • Acyclovir-susceptible isolates exhibited ID50 values ranging from 0.01 to 1.1 µg/ml.
    • The entire assay procedure was completed within 4 hours.

    Conclusions:

    • The developed DNA hybridization method is a simplified, rapid, and objective approach for detecting acyclovir resistance in HSV.
    • This assay facilitates efficient antiviral susceptibility testing, crucial for guiding clinical treatment strategies.
    • The method holds promise for routine use in diagnostic laboratories to manage HSV infections.