Research Project
7035799
DESCRIPTION (provided by applicant): Bioterrorism and biowarfare are a threat for our nation. Such threat became reality in the 2001 Anthrax attack. Although the attack occurred at a small scale, its impact reached terror proportion. Unfortunately, as former Senator Warren Rudman pointed out, "the likelihood of a terrorist attack in this country in the next several years is more likely than unlikely". Recent study by the Council on Foreign Relations concluded that our nation remains "unprepared to handle a catastrophic attack on American soil, particularly one involving chemical, biological or nuclear agents". In the event of a Bioterrorism attack, one of the most important first responses is rapid identification of the biological agent or agents, and timely diagnosis of those who have been infected. Current methods of choice for sensitive detection of B. anthracis are those involving polymerase chain reaction (PCR). The organism is then confirmed with culture methods and/or EIA. The use of PCR is not without problems, however. Possible contamination of PCR amplicons necessitates a specialized and highly controlled central laboratory, which in turn necessitates long distance sample shipping. The fact that PCR is susceptible to inhibitory impurities requires lengthy sample preparation and duplicate testing. All these problems had been encountered during the 2001 Anthrax terror attack. We have developed a rapid (approximately 60 minutes) and sensitive (less than 1 bacterium) prototype sepsis test using our microparticle based amplification (MBA) technology and a novel class of bacterial markers. The specific aims for this Phase I application is to study the feasibility of developing a Bacillus anthracis test system comprising a nucleic acids (pXO1 and pXO2 plasmids) test for the detection of vegetative cells and an agglutination assay for the detection of B. anthracis spores. Both assays use the same MBA technology and are (a) rapid (60-90 minutes), (b) sensitive (< 100 CPU for the nucleic acid test and <5000 spores for the agglutination assay), and (c) specific (little cross reactivity).
