Detection of aerosolized biological agents by immunoassay followed by autonomous PCR confirmation

Dzenitis, John M. ; Hindson, Benjamin J. ; McBride, Mary T. ; Makarewicz, Anthony J. ; Henderer, Bruce D. ; Sathyam, Ujwal S. ; Smith, Sally M. ; Gutierrez, Dora M. ; Metz, Thomas R. ; Venkateswaran, Kodamudi S. ; Colston, Billy W. ; Farrow, Stephen W.

Lawrence Livermore National Laboratory

关键词: Sample Preparation;    Biological Warfare Agents;    Testing;    Aerosol Monitoring;    Lawrence Livermore National Laboratory;   

DOI  :  10.2172/15013780
RP-ID  :  UCRL-TR-202332
RP-ID  :  W-7405-ENG-48
RP-ID  :  15013780

美国|英语

来源: UNT Digital Library

【 摘 要 】

An Autonomous Pathogen Detection System (APDS) unit is an automated, podium-sized system that monitors the air for all three biological threat agents (bacteria, viruses, and toxins). The system has been developed under the auspices of the U. S. Department of Energy and Department of Homeland Security by the University of California, Lawrence Livermore National Laboratory (LLNL) to protect people in critical or high-traffic facilities and at special events. The system performs continuous aerosol collection, sample preparation, and multiplexed biological tests using advanced immunoassays as the primary screen. Over ten agents are assayed at once, and results are reported hourly. R&D work this year focused on incorporating polymerase chain-reaction (PCR) techniques for detecting DNA as confirmation of immunoassay positives. The primary objective of the Dugway testing was to demonstrate the APDS with immunoassay identification and PCR confirmation of bacteria. A secondary objective was to demonstrate immunoassay identification of a protein toxoid (denatured toxin) aerosol release. A total of 12 agent trials were conducted over 14 days of testing, for a total of four work weeks at Dugway. Both testing objectives were achieved with multiple releases and clear identifications. The APDS was shown to be effective for identifying aerosolized Bacillus anthracis, Yersinia pestis, Bacillus globigii, and botulinum toxoid. The two areas for improvement were operational as opposed to hardware-related. The first was slowing the PCR thermal cycling to achieve stronger signals, which was demonstrated during the later phases of testing. The second area is to improve the parameters for autonomous PCR triggering; this is one of the focuses of the upcoming year's work.