A 2002 government study on the accuracy of field tests for anthrax spores casts serious doubt on the validity of these tests. Moreover, one of the DNA markers still being used in some commercial kits was shown to also detect other bacilli commonly found in soil samples that have not been exposed to anthrax. The overall goal of this proposed research is to develop a rapid and specific field assay for early detection of Bacillus anthracis spores using our innovative """"""""Lab-in-a-tube"""""""" (Liat TM) technology. Our Phase I research has established the feasibility of detecting viable anthrax spores and of distinguishing Bacillus anthracis from other Bacillus species. Based on our understanding of the biodefense needs, we will focus on overcoming the challenges of environment sample processing and validating our Liat system for anthrax detection in this Phase II study. The technology developed in this proposed research will ultimately enable the Liat Molecular Analyzer to process both environmental and medical samples and perform rapid real-time PCR detection. We estimate that our live spore assay will be 4-6 times faster and 100-1,000 times more sensitive than the live spore assay currently used.