Martin K. Aune and Nicholas E. Burgis*
Department of Chemistry and Biochemistry, Eastern Washington University, Cheney, WA 99004
Received Date: July 29, 2011; Accepted Date: November 07, 2011; Published Date: November 10, 2011
Citation: Aune MK, Burgis NE (2011) Recent Advances in the Biosciences Aid in the Detection and Identification of Bioterrorism Agents. J Bioterr Biodef S3:e001. doi: 10.4172/2157-2526.S3-e001
Copyright: © 2011 Aune MK, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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The deadly potential of bioterrorism has necessitated the need for warning systems to detect and identify pathogens and toxins used in biological attacks. Biological weapons can be dispersed easily and procured with less difficulty and cost than chemical or nuclear weapons. A major challenge to minimizing outbreaks and casualties is rapid, accurate detection and identification of bioterrorism agents. Development of improved detection and identification methods should be prioritized to insure our national security. Additionally, improved identification methods can aid in forensic evaluation of samples retrieved from the site of attack to help identify the source of the bioterrorism agent used. Recent advancements in detection and identification techniques could prove to be a crucial component in defense against biological attacks.
PCR has seen its use as a common technique for identifying the DNA of pathogens. However, the disadvantage in using classical PCR techniques is that they are time-consuming, and quick analysis is impractical. Recent advancements have produced faster PCR methods which can allow timely diagnosis. Rapid PCR assays make use of accelerated cycling for results within three hours. Use of this technique with enterobacterial species showed accuracy comparable to classical culture techniques in a fraction of the time [1,2]. Real-time PCR assays allow higher specificity and shorter assay times than classical PCR techniques as well. One exciting report describes the development of real-time PCR protocols using reagents that are stable at ambient temperatures. Here the researchers used common real-time PCR techniques to successfully target the F1 antigen gene caf1 in Yersinia pestis. Inclusion of a carbohydrate cocktail and vacuum drying the reagents allowed accurate and reproducible results after six weeks at 37°C . This result is likely to influence the design of portable PCR technologies.
Analysis of field samples procured during law enforcement investigations will also benefit from advances in the biosciences. Advances in electron microscopy have expanded detection limits for viruses and spores. Suspensions of the model bioterrorism agents vaccinia poxvirus and Bacillus subtilis endospores were subjected to negative staining and examination with a 120 kV transmission electron microscope. This technique managed to reduce the detection limit to 105 particles/mL for endospores and to 5 x 104 particles/mL for poxviruses . A crucial factor in accurate pathogen identification is the capability to discriminate between microbial species. Biofilms containing several species can be analyzed using peptide nucleic acid fluorescence in situ hybridization (PNA FISH). This method allows samples to be quantified and visualized while confocal laser scanning microscopy (CLSM) can categorize the biofilm itself . Although more time consuming and labor intensive than the PCR techniques mentioned above, multiplex PCR can combine several PCR assays into one by simultaneously targeting different genes with multiple primers . This can be used in conjunction with real-time PCR by having separate reporter dyes so different signals can be recorded . These techniques are likely to help advance the forensic scientist’s ability to detect and identify bioterrorism agents.
Advanced methodologies have been developed to improve toxin detection as well. A polyclonal antibody microarray using fluorescent nanoparticles has been developed to accurately detect multiplex toxins. This technique was able to detect toxins in beverages and blood at 100 pg/mL . Peptide-functionalized quantum dot resonance energy transfer sensors have been constructed to monitor botulinum toxin activity. The presence of botulinum light-chain protease A can be monitored by permitting self assembly of the protease with an acceptor dye and detection of Förster resonance energy transfer (FRET) . Isotope dilution tandem mass spectrometry can be used to detect ricin toxin in beverages. Ricin is extracted using antibodies attached to magnetic beads. Linear ion traps, used in conjunction with mass spectroscopy, can specifically identify ricin A chain and B chain in test beverages .
Defenses should be prioritized to minimize casualties in biological attacks. Continuing advances in rapid detection of agents used in biological attacks will contribute towards this goal. With greater speed and precision of detection we can better prepare an effective response against biological attacks. Likewise, advances in bioterrorism agent identification can help determine the source of the bioterrorism agent in an effort to bring perpetrators to justice.
The authors research is funded by a grant from the American Heart Association, 10BGIA4370060, awarded to N.E.B.
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