Biothreat agent monitoring using a flow-through polymerase chain reaction instrument.

Extract: The 2001 anthrax letter mailings highlighted critical shortfalls in the USA's capabilities for dealing with the threats of bioterrorism: the lack of effective, reliable, low-cost detection systems for use by state and local authorities. During the few months after the letters were received, 17,000 false alarms and hoaxes were reported, and Americans everywhere were terrified to open their mail. Over 200,000 samples were processed by the Center for Disease Control (CDC) and others. In a scenario where the attack was more wide spread, the laboratory processing requirements could be far greater. In light of such statistics, the imperative for high confidence, high throughput, and inexpensive diagnostics is clear. In answer to this technical challenge, nucleic acid-based methodologies are being standardized and documented to provide acceptable mechanisms of detection. Real time PCR (polymerase chain reaction), in particular, allows analysis of aerosolized or environmental samples to occur within minutes, enabling biological defense response architectures that would otherwise be impossible. PCR is a revolutionary technique for amplifying targeted sequences of DNA. The use of a thermostable enzyme (polymerase) allows sequential dissociation, annealing, and hybridization of complimentary DNA to occur very rapidly. A pair of DNA primers, about 20 nucleotides in length, is used to uniquely identify and amplify (over a million fold) the target DNA sequence.