Thermal Stability of Biothreat Samples Using Solid-Phase FTA® Media

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Authors
  1. Bader, D.E.
  2. Fisher, G.R.
  3. Stratilo, C.W.
Corporate Authors
Defence R&D Canada - Suffield, Ralston ALTA (CAN)
Abstract
FTA® cards were evaluated for their ability to protect PCR amplifiable genetic material from degradation due to extreme and fluctuating temperatures in an effort to assess their thermal protective affects. FTA® cards spotted with purified Bacillus anthracis genomic DNA and live agent were exposed to temperatures ranging from -20ºC to 50ºC and -70ºC to 50ºC, respectively, over a month-long period, and then analyzed using real-time, fluorescence-based PCR assays that detect B. anthracis plasmidic virulence genes. Purified DNA stored on FTA® cards and in solution showed similar thermal stability at low to moderate temperatures but purified DNA stored on FTA® cards was better protected from thermal degradation at very high temperatures (50ºC). In terms of live agent, FTA®-neutralized B. anthracis generated consistent PCR signals at all temperatures compared to agent in solution. Furthermore, a stronger PCR signal was observed for FTA®-neutralized B. anthracis than for live cells and heat-treated cells, even though the FTA®-neutralized PCR reactions contained fewer cells per PCR reaction. The data in this study suggests that FTA® cards offer unique advantages for collecting, storing, and transporting purified DNA or live agent for downstream PCR analysis, particularly in situations where regulated temperature control cannot be guaranteed.

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Report Number
DRDC-SUFFIELD-TM-2009-157 — Technical Memorandum
Date of publication
01 Nov 2009
Number of Pages
48
DSTKIM No
CA036174
CANDIS No
535848
Format(s):
Electronic Document(PDF)

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