Assessment of a Solid Phase Matrix for the Neutralization and Real-Time PCR Detection of Bacillus Anthracis

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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
A commercially available, solid-phase DNA binding matrix (FTA® cards) was evaluated for its ability to neutralize live Bacillus anthracis and entrap nucleic acid for genetic analysis using real-time polymerase chain reaction (PCR) assays. Cell culture analysis of FTA® cards seeded with live B. anthracis indicated that FTA® cards neutralized live B. anthracis but at low concentrations. Therefore, FTA® cards spotted with samples containing, or suspected of containing live B. anthracis should be considered potentially infectious. PCR analysis of FTA® cards seeded with live B. anthracis using assays designed to detect B. anthracis plasmidic gene targets, resulted in detection below the live agent “neutralization” concentration. This may be due in part to a number of factors including multiple plasmids present per colony forming unit (cfu), multiple cells per cfu (cellular clumping), and/or, additional gene target contributions from non-viable cells. PCR reaction solutions exposed to discs seeded with low concentrations of live B. anthracis were found to be culture-negative and thus may be safe to handle under non-containment conditions, but additional studies would be required to determine the level of safety at higher concentrations. Although FTA® cards exhibited limited neutralization capacity for live B. anthracis, they still may be of value for field and lab-based applications as BT/BW agent sample processing, archiving, transport and analysis media, due to their ab

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Keywords
genetic analysis;real-time polymerase chain reaction;solid-phase DNA binding matrix;PCR (Polymerase Chain Reaction)
Report Number
DRDC-SUFFIELD-TM-2006-200 — Technical Memorandum
Date of publication
01 Dec 2006
Number of Pages
32
DSTKIM No
CA029383
CANDIS No
527666
Format(s):
CD ROM

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