title: Stabilization of biothreat diagnostic samples through vitrification matrices.
abstract: Diagnostics for biothreat agents require sample shipment to reference labs for diagnosis of disease; however high/fluctuating temperatures during sample transport negatively affect sample quality and results. Vitrification additives preserve sample integrity for molecular-based assay diagnostics in the absence of refrigeration by imparting whole molecule stability to a plethora of environmental insults. Therefore, we have evaluated commercially available vitrification matrices' (Biomatrica's CloneStable® and RNAStable®) ability to stabilize samples of Yersinia pestis and Venezuelan Equine Encephalitis Virus. When heated to 95°C in RNAStable®, Y. pestis had a 13-fold improvement in detection via real-time PCR compared to heated samples in buffer. VEEV, in RNAStable® at 55°C, had a ~10-fold improved detection versus heated samples in buffer. CloneStable® also preserved Y. pestis antigens for 7days after exposure to cycling temperatures. Overall, RNAStable® and CloneStable® respectively offered superior stabilization to nucleic acids and proteins in response to temperature fluctuations.
authors.affiliation: Diagnostics Systems Division, United States Army Research Institute of Infectious Disease, 1425 Porter Street, Fort Detrick, MD 21702, United States. Electronic address: timothy.d.minogue.civ@mail.mil.
authors.forename: Timothy Devin
authors.initials: TD
authors.lastname: Minogue
authors.affiliation: Diagnostics Systems Division, United States Army Research Institute of Infectious Disease, 1425 Porter Street, Fort Detrick, MD 21702, United States.
authors.forename: Warren Vincent
authors.initials: WV
authors.lastname: Kalina
authors.affiliation: Diagnostics Systems Division, United States Army Research Institute of Infectious Disease, 1425 Porter Street, Fort Detrick, MD 21702, United States.
authors.forename: Susan Rajnik
authors.initials: SR
authors.lastname: Coyne
mesh_terms: D001694:Biological Warfare; D004666:Encephalitis Virus, Venezuelan Equine; D008828:Microbiological Techniques / Q000379:methods*; D009696:Nucleic Acids / Q000032:analysis / Q000737:chemistry; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature; D058989:Vitrification; D015007:Yersinia
chemical_list: D009696:Nucleic Acids
keywords: CloneStable; RNAStable; Venezuelan Equine Encephalitis; Yersina