Abstract: There is an urgent need for a small, inexpensive sensor that can rapidly detect bio-warfare agents with high specificity. Bacillus anthracis, the causative agent of anthrax, would be a perilous disease-causing organism in the event of a release. Currently, most anthrax detection research is based on nucleic acid detection, immunoassays and mass spectrometry, with few detection levels reported below 105 spores. Here, we show the ability to distinguish Bacillus spores to a level approaching 103 spores, below the reported median infectious dose of B. anthracis, using pyrolysis—micromachined differential mobility spectrometry and novel pattern recognition algorithms that combine lead cluster mapping with genetic algorithms. [Copyright &y& Elsevier]