Viral nucleic acid stabilization by RNA extraction reagent.
In the collection of field materials to test for the presence of arboviruses, samples must be appropriately maintained to detect arboviral nucleic acids. In austere field conditions this is often difficult to achieve because, during routine specimen processing, storage, and shipping viral RNA degradation could result in detection failure. RNA extraction reagents, while used commonly for their intended purpose of stabilizing RNA during the extraction process, have not been assessed fully for their potential to stabilize RNA before extraction. The potential for virus stabilization at varying temperatures and periods of time remains unknown. Accordingly, the ability of buffer AVL (Qiagen, Valencia, CA), an RNA extraction reagent, to stabilize viral suspensions of dengue, Venezuelan equine encephalitis and Rift Valley fever viruses was evaluated. The ability of buffer AVL to stabilize each viral suspension was examined at 32, 20, 4, and -20 degrees C. RNA in samples placed in buffer AVL was stable for at least 48h at 32 degrees C and refrigerating samples prolonged stabilization. Additionally, placing the sample/buffer AVL mixture at either 4 or -20 degrees C stabilized samples for at least 35 days. When combined with the ability of buffer AVL to inactivate viral samples, this provides the ability to collect and handle potentially infectious samples in a safe way that also provides sample stabilization.
Blow
Jamie A
JA
Virology Division, United States Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702, USA.
Mores
Christopher N
CN
Dyer
Jessie
J
Dohm
David J
DJ
D001103:Arboviruses / Q000235:genetics / Q000302:isolation & purification*; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification; D004666:Encephalitis Virus, Venezuelan Equine / Q000235:genetics / Q000302:isolation & purification; D005404:Fixatives / Q000378:metabolism*; D011309:Preservation, Biological / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism*; D012296:Rift Valley fever virus / Q000235:genetics / Q000302:isolation & purification; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature; D013997:Time Factors
D005404:Fixatives; D012367:RNA, Viral