Virus inactivation by nucleic acid extraction reagents.
Many assume that common methods to extract viral nucleic acids are able to render a sample non-infectious. It may be that inactivation of infectious virus is incomplete during viral nucleic acid extraction methods. Accordingly, two common viral nucleic acid extraction techniques were evaluated for the ability to inactivate high viral titer specimens. In particular, the potential for TRIzol LS Reagent (Invitrogen Corp., Carlsbad, CA) and AVL Buffer (Qiagen, Valencia, CA) were examined to render suspensions of alphaviruses, flaviviruses, filoviruses and a bunyavirus non-infectious to tissue culture assay. The dilution series for both extraction reagents consistently caused cell death through a 100-fold dilution. Except for the DEN subtype 4 positive control, all viruses had titers of at least 10(6)pfu/ml. No plaques were detected in any extraction reagent plus virus combination in this study, therefore, the extraction reagents appeared to inactivate completely each of the high-titer viruses used in this study. These results support the reliance upon either TRIzol LS Reagent or AVL Buffer to render clinical or environmental samples non-infectious, which has implications for the handling and processing of samples under austere field conditions and low level containment.
Blow
Jamie A
JA
Department 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
Dohm
David J
DJ
Negley
Diane L
DL
Mores
Christopher N
CN
D000524:Alphavirus / Q000187:drug effects; D004279:DNA, Viral / Q000302:isolation & purification*; D005416:Flavivirus / Q000187:drug effects; D007202:Indicators and Reagents / Q000494:pharmacology*; D016851:Orthobunyavirus / Q000187:drug effects; D011933:Reagent Kits, Diagnostic; D014780:Viruses / Q000187:drug effects*
D004279:DNA, Viral; D007202:Indicators and Reagents; D011933:Reagent Kits, Diagnostic