Environmental Monitoring for Biological Threat Agents Using the Autonomous Pathogen Detection System with Multiplexed Polymerase Chain Reaction

Abstract

We have developed and field-tested a now operational civilian biodefense capability that continuously monitors the air in high-risk locations for biological threat agents. This stand-alone instrument, called the Autonomous Pathogen Detection System (APDS), collects and selectively concentrates particles from the air into liquid samples and analyzes the samples using multiplexed PCR amplification coupled with microsphere array detection. During laboratory testing, we evaluated the APDS instrument's response to Bacillus anthracis and Yersinia pestis by spiking the liquid sample stream with viable spores and cells, bead-beaten lysates, and purified DNA extracts. APDS results were also compared to a manual real-time PCR method. Field data acquired during 74 days of continuous operation at a mass-transit subway station are presented to demonstrate the specificity and reliability of the APDS. The U.S. Department of Homeland Security recently selected the APDS reported herein as the first autonomous detector component of their BioWatch antiterrorism program. This sophisticated field-deployed surveillance capability now generates actionable data in one-tenth the time of manual filter collection and analysis.

Biosurveillance: A Review and Update

“…The CDC is responsible for monitoring the reference labs via regular proficiency training [30]. It cannot do the entire job alone and delegates some of the work to states local municipalities.…”

“…This would allow rapid distribution of medical countermeasures, like antibiotics or vaccinations, thereby saving lives [30]. To this end, there are 500 air filters in these 31 urban areas that work as Biowatch sensors.…”

“…These sensors have also been deployed to select indoor venues and are used to monitor mass-gathering events, such as the Super Bowl. This nationwide surveillance system uses distributed aerosol collectors to capture airborne ppapers onto removable dry filters that are transported daily to LRN laboratories for analysis [30]. An expanded deployment of the same technology in 2005 was referred to as Generation 2 Biowatch.…”

Development and Comparison of Two Assay Formats for Parallel Detection of Four Biothreat Pathogens by Using Suspension Microarrays

“…Although high specificity can be achieved by performing careful probe design, differentiation of SNPs based solely on hybridization may be more difficult. Both DH [6], [7], [8], [9] and TSPE-UH formats [10], [11] have been used for multiplex detection of pathogens. While setting up a suspension microarray for the detection of bacterial pathogens, we were confronted with a lack of studies directly comparing the performance of different assay chemistries.…”

Nucleic-Acid Signatures at Three Levels of Readiness for Biowatch

“…Informatics analysis of sequence, including "identification" of strain and sub--strain performed autonomous operation, using multiplexed PCR assays, for the DHS for some period with no false (+) or (--). General operation has been described by Regan, et al 34 .…”