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Virus inactivation by nucleic acid extraction reagents
Jamie A Blow1, David J Dohm, Diane L Negley
1Department of Vector Assessment, United States Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702, USA. jamie.blow@us.army.mil
Journal of Virological Methods
|May 26, 2004
Summary
Common viral nucleic acid extraction methods, TRIzol LS Reagent and AVL Buffer, effectively inactivate high-titer viruses. This ensures sample safety for handling and processing in field conditions and low containment labs.
Area of Science:
- Virology
- Molecular Biology
- Infectious Disease Research
Background:
- Common assumption: viral nucleic acid extraction methods render samples non-infectious.
- Potential for incomplete virus inactivation during extraction processes.
Purpose of the Study:
- Evaluate the inactivation efficacy of two common viral nucleic acid extraction techniques.
- Assess TRIzol LS Reagent and AVL Buffer for inactivating high-titer alphaviruses, flaviviruses, filoviruses, and bunyaviruses.
Main Methods:
- Tested TRIzol LS Reagent and AVL Buffer against high-titer virus suspensions.
- Utilized tissue culture assays to detect viral infectivity post-treatment.
- Included a dilution series to determine the effective inactivation concentration.
Main Results:
- Both extraction reagents caused cell death through a 100-fold dilution.
- All tested viruses, except one subtype, had initial titers of at least 10^6 plaque-forming units/ml.
- No viral plaques were detected in any virus-reagent combination, indicating complete inactivation.
Conclusions:
- TRIzol LS Reagent and AVL Buffer effectively inactivate high-titer viruses.
- These reagents can be relied upon to render clinical and environmental samples non-infectious.
- Supports safe handling and processing of samples under austere field conditions and low containment.