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Detection of biological warfare agents with fiber-optic microsphere-based DNA arrays

Biological warfare agents (BWAs) pose significant threats to both military forces and civilian populations. The increased concern about bioterrorism has promoted the development of rapid, sensitive, and reliable detection systems to provide an early warning for detecting the release of BWAs. We have developed a high-density DNA array to detect BWAs in real environmental samples with fast response times and high sensitivity. An optical fiber bundle containing approximately 50,000 individual 3.1 μm diameter fibers was chemically etched to yield an array of microwells and used as the substrate for the array. 50-mer single-stranded DNA probes designed to be specific for target BWAs were covalently attached to 3.1-μm microspheres, and the microspheres were distributed into the microwells to form a randomized high-density DNA array. We demonstrated the applicability of this DNA array for the identification of Bacillus thuringiensis kurstaki, a BWA simulant, in real samples. PCR was used to amplify the sequences, introduce fluorescent labels into the target molecules, and provide a second level of specificity. After hybridization of test solutions to the array, analysis was performed by evaluating the specific responses of individual probes on the array.