Stabilization of biothreat diagnostic samples through vitrification matrices

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Biothreat agent diagnostics require sample transport, which is often compromised by high or fluctuating temperatures.
• Temperature extremes negatively impact sample quality and the accuracy of diagnostic results.
• Preserving sample integrity during transport is crucial for reliable disease diagnosis.

Purpose of the Study:

• To evaluate the efficacy of commercially available vitrification matrices (CloneStable® and RNAStable®) in stabilizing biothreat agent samples.
• To assess the impact of these matrices on the stability of nucleic acids and proteins under thermal stress.
• To determine the improvement in diagnostic detection rates when using vitrification additives.

Main Methods:

• Samples of Yersinia pestis (bacteria) and Venezuelan Equine Encephalitis Virus (VEEV) were stabilized using RNAStable® and CloneStable®.
• Stabilized and unstabilized samples were subjected to elevated temperatures (95°C for Y. pestis, 55°C for VEEV) and cycling temperatures.
• Sample integrity and detection rates were assessed using real-time PCR for nucleic acids and antigen detection methods for proteins.

Main Results:

• RNAStable® significantly improved the detection of Y. pestis by 13-fold via real-time PCR after heating to 95°C compared to controls.
• RNAStable® enhanced VEEV detection by approximately 10-fold at 55°C compared to samples in buffer.
• CloneStable® preserved Y. pestis antigens for 7 days under cycling temperature conditions.

Conclusions:

• RNAStable® and CloneStable® provide superior stabilization for nucleic acids and proteins, respectively, against temperature fluctuations.
• Vitrification additives are effective in maintaining sample integrity for molecular-based diagnostics in challenging transport conditions.
• These findings support the use of vitrification matrices to improve the reliability of biothreat agent diagnostics.