Real-Time Detection of Biological Pathogens in Urban Environments Laura Barry Hung Phan Gloria See Introduction to Biosensors Presentation 3 - 5/10/2011

Project Details Pathogen are agents known to cause disease in living organisms such as plants, animals, and humans Pathogens may include: Bacteria Viruses Biotoxins Fungus A review of biosensor technology for real-time pathogen detection in urban environments.

Traditional Methods of Detection & Identification Methods require time and labor intensive stages: Sample w/ unidentified agent: air, soil, food/water, sputum etc. Purification Culture enhancement Selective/differential plating Isolation and subculturing Inoculation for physiological and biochemical tests Evaluation of physiological and biochemical tests sciencedude.ocregister.com gene-quantification.de

Basic components of sensor system

Advances in biosensor technology Approaches Point detection Standoff technologies Passive standoff technologies Lab-on-a-chip Technologies Cell-based Structure based identification Nucleic acid sequence identification Light scattering Optical Infrared Two-component reporter system Microfluidics

Current Biosensor Technologies Diagnostic Assays o Uses probes and assays to identify Pathogen o Identifies known select genome strands in pathogen to detect families of pathogen o Identifies full strands of known genome to identify specific pathogen o Sensitive o Fast o Expensive

Current Biosensor Technologies Microfluidics o Tiny chips that have fluid channels and sensing chambers o Lab-on-chip o Uses nucleic acid-base to detect pathogens  E-noses  Can identify single molecule in samples  Can flag DNA strands o Ergonomic, Fast, Expensive

Current Biosensor Technologies Optical Sensors o Fluorescence  Optical multisensors, usually with each specialized with a single reactant  organic materials react with the sensing surface and change the intensity of the light o Surface Plasmon Resonance  target is collected  object’s surface is excited by a light source and it oscillates to a certain frequency

Current Biosensor Technologies Bioreporters o using cells or other organisms to detect relevant stimuli o utilizes inherently rapid response times of cellular signaling o easy to use o reporting system needs to be established in advance o organisms need to be maintained in their testing environment Bomb-Sniffing Plants Colorado State University

Current Biosensor Technologies Other approaches o Electrochemical: Used for DNA and immunusensors o Electromechanical: Usually mass-change sensors that attract pathogens o Chemical: Detecting specific compounds native to specimen, usually utilizing surface chemistry

Current Biosensor Technologies Nanomaterials o Coupled with other sensing technologies, nanomaterials expand the potential for biosensors o Cell and molecule scale features make detection easier o Sensitivity and threshold limits are improved o Increased surface area gives more room for substrates to be treated, detection and increases device sensitivity

Relevant Applications: Airport, subways, high human traffic environments (DHS, TSA) Transportation Security Administration Borders, ports (DHS, DOS) Battlefield Infrastructure (WHO, EPA, industry, National Biodefense Program) Emergency Responders

Existing Systems – E-Noses: Inspired from nature uses a combination of new technologies to detect pathogens reduces time from days to fifteen minute intervals currently being developed to detect spoiled meat NASA's E-Nose courtesy JPL courtesy of 4to40.com

Existing Systems - BioWatch Program: Department of Homeland Security (DHS) Pathogen sensors are mounted at air quality monitoring stations. Sensors collects particles in the air which pass through filters. Based on BASIS, (Biological Aerosol Sentry and Information System). Design of filter mechanism uses automatic sequential filtering. Deployed in 2002 for indoor/outdoor monitoring at the Olympics, tested for urban environment. Only a few of the results from these tests were released, it was shown to have high specificity and sensitivity while having less than 0.005% false positives. The system is labor intensive. October 2003, Texas - first positive result of tularemia was reported, detection was modest, precautionary measures were taken

Existing Systems – TB Breathalyzer Rapid Biosensor Systems is a UK based development company Technology for rapid screening of infectious agents. TB Breathalyser device for screening within a few minutes. Analyses the sample by via displacement assay utilizing the evanescent wave and bio-optical sensing technologies Design: small, robust, simple to use, non-invasive and low cost. January 2011: entered into agreement with Ortho Clinical Diagnostics, Inc. to bring the breathalyser into production.

Future Challenges in Biosensor Technology Detection limits/sensitivity Multisensor devices Reliability/accuracy of detection Speed of analysis Field environments Reusable, robust devices Cost of fabrication and testing

References Friedli, Andrienne. Phase 1 Final Report. SERRI Project: Biosensor Research. Supported by Dept. Homeland Security & Dept. of Energy Interagency Agreement. Introduction to Biological Agent Detection Equipment for Emergency First Responders, NIJ Guide NCJ , Alim A. Fatah; John A. Barrett; Richard D. Arcilesi Jr.; Kenneth J. Ewing; Charlotte H. Lattin; Timothy F. Moshier, December 2001, NIJ, (53 pages). Donaldson, Kim A. et. al. A rapid detection method for Vaccinia virus, the surrogate for smallpox virus Jinseok Heo, et. al. An Overview of Recent Strategies in Pathogen Sensing Cynthia J. Bruckner-Lea. Biosensor Systems for Homeland Security Biosensors for Anthrax Detection - Program Overview From Counter Terrorism to Pathogen Detections Multiplexed Electrochemical Detection of Yersinia Pestis and... Nanosensors in the Age of Terror New Biosensor for Rapid Detection of Anthrax Lethal Toxin Nucleic Acid-based Detection of Bacterial Pathogens... Pathogen Biosensors Rapid detection of Bacillus anthracis...

References The Autonomous Pathogen Detection System Environmental Microbiology: Current Technology and Water Applications (Keya Sengupta) 2011 Chapter 12: The Microbe as a Reporter: Microbial Bioreporter Sensing Technologies for Chemical and Biological Detection (pg 281) Ripp. Layton, Sayler ( eClEls7TAz&sig=OwBzdesm2DNtiB_NcNp5kS84LyY#v=onepage&q=urban%20detection%20biological%20pathogens&f=false) eClEls7TAz&sig=OwBzdesm2DNtiB_NcNp5kS84LyY#v=onepage&q=urban%20detection%20biological%20pathogens&f= The Coming Together of the Sciences: Biosensors for the Detection of Waterborne Pathogens Using Antibodeies and Gene-based Recognition Chemistries (Sen Xu and Raj Mutharasan) Microbial Source Tracking (Domingo, Lamendella, Ashbolt) Chemical and Biological Weapons: Current Concepts for Future Defenses Plamen A. Demirev, Andrew B. Feldman, and Jeffrey S. Lin. Johns Hopkins APL Technical Digest, Volume 26, Number A high-throughput pipeline for designing mircroarray-based pathogen diagnostic assays. Ravi Vijaya Satya, Nela Zavaljevski, Kamal Kumar and Jaques Reifman. BMC Bioinformatics 2008, 9:185. Published 10 April Antunes MS, Morey KJ, Smith JJ, Albrecht KD, Bowen TA, et al. (2011) Programmable Ligand Detection System in Plants through a Synthetic Signal Transduction Pathway. PLoS ONE 6(1): e doi: /journal.pone An Optofluidic Nanoplasmonic Biosensor for Direct Detection of Live Viruses from Biological Media Ahmet A. Yanik, Min Huang, Osami Kamohara, Alp Artar, Thomas W. Geisbert, John H. Connor, Hatice Altug Nano Letters (12),

References Nucleic Acid Amplification Strategies for DNA Microarray-Based Pathogen Detection Gary J. Vora,1* Carolyn E. Meador,2 David A. Stenger,1 and Joanne D. Andreadis1† Feng, Peter. Rapid Methods for Detecting Foodborne Pathogens. Bacteriological Analytical Manual, 8th Edition, January Zourob, M. et al. ed. Principles of Bacterial Detection: Biosensors, Recognition Receptors and Microsystems. Springer, New York, NY, 2008 Sack, Kevin and Timothy Williams. “Deaths of 9 Alabama Patients Tied to Intravenous Supplement”. The New York Times [New York City] 31 March 2011, page A20. Shea, Dana A. and Sarah A. Lister. “The BioWatch Program: Detection of Bioterrorism”. Congressional Research Service Report No. RL November 19, 2003 Camilleri, Dennis. “New Screening Solution Offers Hope in the Battle Against TB”. Rapid Biosensor Systems