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Stabilization of biothreat diagnostic samples through vitrification matrices

J Microbiol Methods. 2014 Jun:101:81-5. doi: 10.1016/j.mimet.2014.04.002. Epub 2014 Apr 15.

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.

Keywords: CloneStable; RNAStable; Venezuelan Equine Encephalitis; Yersina.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Warfare*
  • Encephalitis Virus, Venezuelan Equine
  • Microbiological Techniques / methods*
  • Nucleic Acids / analysis
  • Nucleic Acids / chemistry
  • Specimen Handling / methods*
  • Temperature
  • Vitrification*
  • Yersinia

Substances

  • Nucleic Acids