Herpes Simplex and Varicella Zoster Viruses

Questions and Answers

December 2009

Another question that you may have already thought of is whether or not there is an advantage to using M5 media for maintaining detectable DNA during transport. Clearly, we wouldn’t want to recommend the culturette tube for transport if the M5 media provided a significant advantage.

Well, it turns out that M5, while an excellent transport media for viral culture, is not actually beneficial for PCR detection of HSV and VZV from dermal and anogenital swabs. In fact, it is better to submit the sample in a culturette tube, because the virus does not need to be viable in order for DNA to be detected, and the addition of M5 media may dilute the sample, and decrease the sensitivity of detection.

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Jump to section:

• Introduction
• The Viruses
• Laboratory Diagnosis
• HSV/VZV Real-time PCR Sample Workflow
• Is Extraction Required for Dermal and Anogenital Specimens?
• Direct Processing in Neutralization Buffer: An Alternative to MagNA Pure Extraction
• Match Up Identifying Information
• Remove Swab from the Culturette Tube
• Heat Swab in Neutralization Buffer
• Sample is Ready for PCR
• A New Workflow for Dermal and Anogenital Swabs
• Direct Processing in Neutralization Buffer: Comparable to DNA Extraction?
• Comparison of Detection - HSV
• Results: Comparison of Amplification Cycle threshold
• Comparison of Detection - VZV
• Results: Comparison of Amplification Crossing Points
• Questions and Answers
• Questions and Answers
• M5 Media vs Culturette Tubes: Experiment
• M5 Media vs Culturette Tubes: Results
• Conclusions
• Questions?