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

Abstract

A commercially available, solid-phase DNA binding matrix (FTA (Trademark) 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 (Trademark) cards seeded with live B. anthracis indicated that FTA (Trademark) cards neutralized live B. anthracis but at low concentrations. Therefore, FTA (Trademark) cards spotted with samples containing, or suspected of containing live B. anthracis should be considered potentially infectious. PCR analysis of FTA (Trademark) 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 (Trademark) 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 ability to protect and preserve genetic material under storage or transport conditions where cryopreservation (refrigeration or freezing) may not be possible and, therefore, are of continued interest.

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Document Details

Document Type
Technical Report
Publication Date
Dec 01, 2006
Accession Number
ADA472833

Entities

People

  • Chad W. Stratilo
  • Douglas E. Bader
  • Glen R. Fisher

Tags

DTIC Thesaurus Topics

  • Acids
  • Cell Count
  • Chain Reactions
  • Chemical Reactions
  • Classification
  • Culture Techniques
  • Deoxyribonucleic Acids
  • Detection
  • Materials
  • National Security
  • Neutralization
  • Nucleic Acids
  • Pcr Testing
  • Phase
  • Polymerase Chain Reaction
  • Security
  • Solid Phases

Fields of Study

  • Biology

Readers

  • Immunology
  • Microbial Pathology
  • Molecular Genetics

Technology Areas

  • Biotechnology