Real-Time Detection of Biological Pathogens in Urban

Environments

Laura BarryHung PhanGloria See

Introduction to BiosensorsPresentation 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:BacteriaVirusesBiotoxinsFungus

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 Assayso Uses probes and assays to identify Pathogeno Identifies known select genome strands in pathogen

to detect families of pathogeno Identifies full strands of known genome to identify

specific pathogeno Sensitiveo Fasto Expensive

Current Biosensor Technologies• Microfluidics

o Tiny chips that have fluid channels and sensing chambers

o Lab-on-chipo Uses nucleic acid-base to

detect pathogens E-noses Can identify single molecule

in samples Can flag DNA strands

o Ergonomic, Fast, Expensive

http://www.indiatalkies.com

http://futuremed2011.com/headlines-from-the-future/

Current Biosensor Technologies

• Optical Sensorso 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 stimulio utilizes inherently rapid response times of cellular signalingo easy to useo reporting system needs to be established in advanceo organisms need to be maintained in their testing

environment Bomb-Sniffing Plants Colorado State University

Current Biosensor Technologies

• Other approacheso Electrochemical: Used for DNA and immunusensorso Electromechanical: Usually mass-change sensors that

attract pathogenso Chemical: Detecting specific compounds native to

specimen, usually utilizing surface chemistry

Current Biosensor Technologies

• Nanomaterialso Coupled with other sensing technologies, nanomaterials

expand the potential for biosensorso Cell and molecule scale features make detection easiero Sensitivity and threshold limits are improvedo 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

ReferencesFriedli, 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 101-00. NCJ 190747, 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...

ReferencesThe 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 (http://books.google.com/books?hl=en&lr=&id=1gjZxeeqil4C&oi=fnd&pg=PA281&dq=urban+detection+biological+pathogens&ots=eClEls7TAz&sig=OwBzdesm2DNtiB_NcNp5kS84LyY#v=onepage&q=urban%20detection%20biological%20pathogens&f=false)

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 4. 2005.

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 2008.

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): e16292. doi:10.1371/journal.pone.0016292

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 2010 10 (12), 4962-4969

ReferencesNucleic 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 2001.

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 32152. November 19, 2003

Camilleri, Dennis. “New Screening Solution Offers Hope in the Battle Against TB”. Rapid Biosensor Systems. 2008. http://www.rapidbiosensor.com/pdf/RBS%20New%20Screening%20Solution.pdf