U.S. Department of Justice

Office of Justice Programs

National Institute of Justice

Law Enforcement and Corrections Standards and Testing Program

National Institute of Justice

An Introduction to Biological Agent 

Detection Equipment for 

Emergency First Responders

NIJ Guide 101–00

December 2001
U.S. Department of Justice

Office of Justice Programs

810 Seventh Street N.W.

Washington, DC 20531

John Ashcroft

Attorney General

Deborah J. Daniels

Assistant Attorney General

Sarah V. Hart

Director, National Institute of Justice

For grant and funding information, contact:

Department of Justice Response Center 

800–421–6770

Office of Justice Programs National Institute of Justice

World Wide Web Site World Wide Web Site 

http://www.ojp.usdoj.gov http://www.ojp.usdoj.gov/nij
U.S. Department of Justice

Office of Justice Programs

National Institute of Justice

An Introduction to Biological Agent Detection Equipment for

Emergency First Responders

NIJ Guide 101-00

Dr. Alim A. Fatah1

John A. Barrett2

Richard D. Arcilesi, Jr.2

Dr. Kenneth J. Ewing2

Charlotte H. Lattin2

LTC Timothy F. Moshier3

Coordination by:

Office of Law Enforcement Standards

National Institute of Standards and Technology

Gaithersburg, MD 20899

Prepared for:

National Institute of Justice

Office of Science and Technology

Washington, DC 20531

December 2001

 NCJ 190747

 

1National Institute of Standards and Technology, Office of Law Enforcement Standards.

2Battelle Memorial Institute.

3

Joint Program Office for Biological Defense (JPOBD).
National Institute of Justice

Sarah V. Hart

Director

This guide was prepared for the National Institute of Justice, U.S. Department of Justice, by the Office of Law

Enforcement Standards of the National Institute of Standards and Technology under Interagency Agreement

94–IJ–R–004, Project No. 99–060–CBW. It was also prepared under CBIAC contract No. SPO–900–94–D–0002

and Interagency Agreement M92361 between NIST and the Department of Defense Technical Information Center

(DTIC).

The authors wish to thank Ms. Kathleen Higgins of the National Institute of Standards and Technology, Mr. Bill

Haskell of SBCCOM, Ms. Priscilla S. Golden of General Physics, LTC Don Buley of the Joint Program Office of

Biological Defense, Ms. Nicole Trudel of Camber Corporation, Dr. Stephen Morse of Centers for Disease Control,

and Mr. Todd Brethauer of the Technical Support Working Group for their significant contributions to this effort.

We would also like to acknowledge the Interagency Board for Equipment Standardization and Interoperability,

which consists of Government and first responder representatives.
iii

FOREWORD

The Office of Law Enforcement Standards (OLES) of the National Institute of Standards and

Technology (NIST) furnishes technical support to the National Institute of Justice (NIJ) program to

support law enforcement and criminal justice in the United States. OLES’s function is to develop

standards and conduct research that will assist law enforcement and criminal justice agencies.

OLES is: (1) subjecting existing equipment to laboratory testing and evaluation, and (2) conducting

research leading to the development of several series of documents, including national standards,

user guides, and technical reports.

This document covers research conducted by OLES under the sponsorship of NIJ. Additional

reports as well as other documents are being issued under the OLES program in the areas of

protective clothing and equipment, communications systems, emergency equipment, investigative

aids, security systems, vehicles, weapons, and analytical techniques and standard reference

materials used by the forensic community.

Technical comments and suggestions concerning this guide are invited from all interested parties.

They may be addressed to the Office of Law Enforcement Standards, National Institute of Standards

and Technology, 100 Bureau Drive, Stop 8102, Gaithersburg, MD 20899−8102.

 Sarah V. Hart, Director

 National Institute of Justice
v

CONTENTS

FOREWORD..................................................................................................................................iii

COMMONLY USED SYMBOLS AND ABBREVIATIONS .....................................................vii

ABOUT THIS GUIDE...................................................................................................................ix

1. INTRODUCTION..................................................................................................................3

2. REVIEW OF BIOLOGICAL AGENTS.................................................................................5

2.1 Bacterial Agents............................................................................................................5

2.2 Viral Agents...................................................................................................................5

2.3 Rickettsiae......................................................................................................................5

2.4 Biological Toxins...........................................................................................................5

3. CHALLENGES TO BIOLOGICAL AGENT DETECTION ..............................................13

3.1 The Ambient Environment...........................................................................................13

3.2 Selectivity of the Detection System.............................................................................15

3.3 Sensitivity.....................................................................................................................15

3.4 Sampling.......................................................................................................................15

4. BIOLOGICAL DETECTION SYSTEM COMPONENTS..................................................17

4.1 Trigger/Cue ..................................................................................................................17

4.2 Collector.......................................................................................................................18

4.3 Detector........................................................................................................................18

4.4 Identifier.......................................................................................................................18

5. OVERVIEW OF BIOLOGICAL AGENT DETECTION SYSTEM TECHNOLOGIES ...19

5.1 Point Detection Technologies......................................................................................20

5.2 Standoff Technologies.................................................................................................33

5.3 Passive Standoff Technologies....................................................................................35

6. HOW TO PREPARE FOR A BIOLOGICAL INCIDENT... ..............................................37

6.1 Federal and State Programs for Support......................................................................37

6.2 Crisis Management in a Terrorist Attack.....................................................................38

6.3 Functional Tasks During a Terrorist Attack ................................................................38

7. SUMMARY.........................................................................................................................41

APPENDIX A––REFERENCES................................................................................................A−1

APPENDIX B––CONTACT INFORMATION FOR FIRST RESPONDERS..........................B−1

TABLES

Table 2−1. Bacterial agents...........................................................................................................7

Table 2−2. Viral agents.................................................................................................................9

Table 2−3. Ricksettsiae ...............................................................................................................11

Table 2−4. Biological toxins.......................................................................................................12

FIGURES

Figure 1. Comparative toxicity of effective doses of biological agents, toxins, and

chemical agents...........................................................................................................1
vi

Figure 3−1. Airborne bacterial concentration fluctuation in a single day...................................14

Figure 4−1. Typical point detection automated architecture (with a combined trigger/cue)......17

Figure 5−1. Biological Integrated Detection System (BIDS).....................................................20

Figure 5−2. Cutaway of UK Integrated Biological Detection System (IBDS)...........................20

Figure 5−3. FLAPS II (component of the Canadian 4WARN System)......................................22

Figure 5−4. Canadian Integrated Biological-Chemical Agent Detection System

(CIBADS)/4WARN ................................................................................................22

Figure 5−5. BioVICTM Aerosol Collector, MesoSystems Technology, Inc................................24

Figure 5−6. Joint Biological Point Detection System (JBPDS)..................................................25

Figure 5−7. Smart Air Sampler System (SASS 2000), Research International..........................25

Figure 5−8. BioCaptureTM BT-500 Air Sampler, MesoSystems Technology, Inc......................26

Figure 5−9. B-D Flow Cytometer FACSCaliber, Becton Dickenson.........................................28

Figure 5−10. Chemical Biological Mass Spectrometer (CBMS), Bruker....................................29

Figure 5−11. BTATM Test Strip testing procedure, Tetracore, LCC .............................................30

Figure 5−12. NDI Smart Ticket....................................................................................................31

Figure 5−13. Rapid LightCycler, Idaho Technology.................................................................33

Figure 5−14. RAPID, Idaho Technology......................................................................................33

Figure 5−15. Long-Range Biological Standoff Detection System (LIDARS).............................35
vii

COMMONLY USED SYMBOLS AND ABBREVIATIONS

A ampere hf high frequency o.d. outside diameter

ac alternating current Hz hertz Ω ohm

AM amplitude modulation i.d. inside diameter p. page

cd candela in inch Pa pascal

cm centimeter IR infrared pe probable error

CP chemically pure J joule pp. pages

c/s cycle per second L lambert ppm parts per million

d day L liter qt quart

dB decibel lb pound rad radian

dc direct current lbf pound-force rf radio frequency

°C degree Celsius lbf†in pound-force inch rh relative humidity

°F degree Fahrenheit lm lumen s second

dia diameter ln logarithm (base e) SD standard deviation

emf electromotive force log logarithm (base 10) sec. Section

eq equation M molar SWR standing wave ratio

F farad m meter uhf ultrahigh frequency

fc footcandle μ micron UV ultraviolet

fig. Figure min minute V volt

FM frequency modulation mm millimeter vhf very high frequency

ft foot mph miles per hour W watt

ft/s foot per second m/s meter per second λ wavelength

g acceleration mo month wk week

g gram N newton wt weight

gr grain N†m newton meter yr year

H henry nm nanometer

h hour No. number

area=unit2 (e.g., ft2, in2, etc.); volume=unit3 (e.g., ft3, m3, etc.)

ACRONYMS SPECIFIC TO THIS DOCUMENT

APS Aerosol Particle Sizer IND Investigational New Drug

BA Biological Agent IR Infrared

BAWS Biological Aerosol Warning System JSLSCAD Joint Service Lightweight Standoff Chemical

Agent Detector

BDG Bi-Diffractive Grating LANL Los Alamos National Laboratory

BW Biological Warfare LD50 Lethal Dose for 50% of Population

CA Chemical Agent LIDAR Light Detection and Ranging

CBMS Chemical Biological Mass Spectrometer LLNL Lawrence Livermore National Laboratory

CIBADS Canadian Integrated Biological Agent Detection

System

MALDI-TOF Matrix Assisted Laser Desorption IonizationTime of Flight

CW Chemical Warfare mg Milligram

DARPA Defense Advanced Research Projects Agency NASA National Aeronautical Space Administration

DNA Deoxyribonucleic Acid PCR Polymerase Chain Reaction

DoD BSK Department of Defense Biological Sampling Kit PHTLAAS Portable High-Throughput Liquid Aerosol Air

Sampler System

DOE Department of Energy PY-GC-IMS Pyrolysis-Gas Chromatography-Ion Mobility

Specrometer

ECBC Edgewood Chemical and Biological Command RNA Ribonucleic Acid

EOO Electro Optics Organization, Inc. RSCAAL Remote Sensing Chemical Agent Alarm

FLAPS Fluorescent Aerodynamic Particle Sizer SBCCOM Soldier and Biological Chemical Command

FTIR Fourier Transform Infrared SESI Science and Engineering Services, Inc.

HHA Hand-Held Immunochromatographic Assay SRI Stanford Research Institute

HeNe Helium-Neon TE Transverse Electric

HUS Hemolytic uremic syndrome TIMs Toxic Industrial Materials

HVAPS High Volume Aerodynamic Particle Sizer TM Transverse Magnetic

IAB Interagency Board TTP Thrombocytopenic purpura

IBADS Interim Biological Agent Detector System UAV Unmanned Aerial Vehicle

IMS Ionization/Ion Mobility Spectrometry WMD Weapons of Mass Destruction
viii

PREFIXES (See ASTM E380) COMMON CONVERSIONS

d deci (10-1) da deka (10) 0.30480 m =1 ft 4.448222 N = 1 lbf

c centi (10-2) h hecto (102) 25.4 mm = 1 in 1.355818 J = 1 ft†lbf

m milli (10-3) k kilo (103) 0.4535924 kg = 1 lb 0.1129848 N m = 1 lbf†in

μ micro (10-6) M mega (106) 0.06479891g = 1gr 14.59390 N/m = 1 lbf/ft

n nano (10-9) G giga (109) 0.9463529 L = 1 qt 6894.757 Pa = 1 lbf/in2

p pico (10-12) T tera (1012) 3600000 J = 1 kW†hr 1.609344 km/h = 1 mph

psi = mm of Hg x (1.9339 x 10-2)

mm of Hg = psi x 51.71

Temperature: T °C = (T °F –32)×5/9 Temperature: T °F = (T °C ×9/5)+32
ix

ABOUT THIS GUIDE

The National Institute of Justice (NIJ) is the focal point for providing support to State and local law

enforcement agencies in the development of counterterrorism technology and standards, including

technological needs for chemical and biological defense. In recognizing the needs of State and local

emergency first responders, the Office of Law Enforcement Standards (OLES) at the National

Institute of Standards and Technology (NIST), working with NIJ, the Technical Support Working

Group (TSWG), the U.S. Army Soldier and Biological Chemical Command (SBCCOM), and the

Interagency Board for Equipment Standardization and Interoperability (IAB), is developing

chemical and biological defense equipment guides. The guides will focus on chemical and

biological equipment in areas of detection, personal protection, decontamination, and

communication. This document focuses specifically on assisting the emergency first responder

community in the understanding of biological agent detection equipment.

The long range plans are to: (1) subject existing biological agent detection equipment to laboratory

testing and evaluation against a specified protocol, and (2) conduct research leading to the

development of multiple series of documents, including national standards, user guides, and

technical reports. It is anticipated that the testing, evaluation, and research processes will take

several years to complete; therefore, NIJ has developed this initial guide for the emergency first

responder community in order to facilitate an understanding of biological agent detection equipment.

In conjunction with this program, additional guides, as well as other documents, are being issued in

the areas of chemical agent and toxic industrial material detection equipment, decontamination

equipment, personal protective equipment, and communications equipment used in conjunction with

protective clothing and respiratory equipment.

The information contained in this guide on specific equipment and technologies has been obtained

through literature searches and market surveys. Reference herein to any specific commercial

products, processes, or services by trade name, trademark, manufacturer, or otherwise does not

necessarily constitute or imply its endorsement, recommendation, or favoring by the United States

Government. The information and statements contained in this guide shall not be used for the

purposes of advertising, nor to imply the endorsement or recommendation of the United States

Government.

With respect to information provided in this guide, neither the United States Government nor any of

its employees make any warranty, expressed or implied, including but not limited to the warranties

of merchantability and fitness for a particular purpose. Further, neither the United States

Government nor any of its employees assume any legal liability or responsibility for the accuracy,

completeness, or usefulness of any information, apparatus, product, or process disclosed.

Technical comments, suggestions, and product updates are encouraged from interested parties. They

may be addressed to the Office of Law Enforcement Standards, National Institute of Standards and

Technology, 100 Bureau Drive, Stop 8102, Gaithersburg, MD 20899−8102. It is anticipated that

this guide will be updated periodically.
1

AN INTRODUCTION TO BIOLOGICAL AGENT DETECTION

EQUIPMENT FOR EMERGENCY FIRST RESPONDERS

The end of the cold war has reduced international tension between the super powers. However,

ironically enough, this has resulted in regional instability due to a resurgence of nationalistic,

religious, and ethnic strife, which presents a real threat to peace in all regions of the globe.

Additionally, there has been a remarkable increase in the production and availability of chemical

and biological weapons throughout the world. The combination of these factors has significantly

increased the possibility of an attack on the United States involving the use of such weapons.

Biological agents are often considered to be psychologically the more threatening of the two, and

therefore provide more appeal to the terrorist.

Biological agents can be manufactured in facilities that are inexpensive to construct; that

resemble pharmaceutical, food, or medical production sites; and that provide no detectable sign

that such agents are being produced. One characteristic of biological agents that makes them so

attractive to potential users is their remarkably low effective dose; that is, the mass of agent that

is required to create the desired effect (incapacitation or death) on the target population. Figure 1

shows the approximate mass in milligrams (mg) of an agent needed to achieve the desired result

in comparison to toxins and chemical agents. The mass of a paper clip is included in this

diagram as a point of reference. The reader can immediately see the vast differences in

effectiveness between biological agents (microbial agents, e.g., bacteria and viruses) and

chemical agents. At the extreme, some biological agents are as much as 14 billion times more

effective than chemical agents, making it easy to see why biological agents are often described as

the poor man’s atomic bomb. The reader should also note that if a terrorist chooses to use a

toxin agent (in order to get relatively rapid effects in a tactical situation), a much greater mass of

the toxin agent will have to be employed than if biological agents were being used. This mass of

toxin agent in some cases may be equivalent to chemical agent masses.

1 paper clip weighs

about 500 mg

1mg

BW Agents (Pathogens)

.0000001 .000001 .00001 .0001 .001 .01 .1 10 100 1000

Toxin Agents

CW Agents

10000

Figure 1. Comparative toxicity of effective doses of biological agents, toxins,

and chemical agents
3

1. INTRODUCTION

The primary purpose of this document is to function as a guide and provide emergency first

responders with information to aid them in their understanding of biological agent detection

equipment.

This document is divided into seven sections and includes two appendices. Section 2 presents a

review of biological agents. Specifically, it discusses the four most common classes of

biological agents and provides information that includes epidemiology, symptoms, and

treatment. Section 3 provides an overview of the known challenges associated with biological

agent detection. Specifically, this section discusses general detection requirements such as

ambient environment, selectivity, sensitivity, and sampling. Section 4 provides the reader with

background information on the components of biological detection systems. Section 5 discusses

known detection technologies, identified as point, standoff, or active standoff detection. Section

6 provides the emergency first responder with information on how to prepare for a biological

incident. Section 7 concludes by providing a concise summary of the current state of biological

agent detection. Appendix A identifies the sources of information used in developing this

document. Appendix B provides contact information (telephone numbers and internet addresses)

for State public health laboratories.
5

2. REVIEW OF BIOLOGICAL AGENTS

This section provides a description of the biological agents likely to be used in a terrorist attack.

There are four categories under discussion: bacterial agents (sec. 2.1), viral agents (sec. 2.2),

rickettsiae (sec. 2.3), and biological toxins (sec. 2.4).

2.1 Bacterial Agents

Bacteria are small, single-celled organisms, most of which can be grown on solid or in liquid

culture media. Under special circumstances, some types of bacteria can transform into spores

that are more resistant to cold, heat, drying, chemicals, and radiation than the bacterium itself.

Most bacteria do not cause disease in human beings, but those that do cause disease act in two

differing mechanisms: by invading the tissues or by producing poisons (toxins). Many bacteria,

such as anthrax, have properties that make them attractive as potential warfare agents:

• Retained potency during growth and processing to the end product (biological weapon).

• Long “shelf-life.”

• Low biological decay as an aerosol.

Other bacteria require stabilizers to improve their potential for use as biological weapons. Table

2−1 lists some of the common bacterial agents along with possible methods of dissemination,

incubation period, symptoms, and treatment.

2.2 Viral Agents

Viruses are the simplest type of microorganism and consist of a nucleocapsid protein coat

containing genetic material, either RNA or DNA. Because viruses lack a system for their own

metabolism, they require living hosts (cells of an infected organism) for replication. As

biological agents, they are attractive because many do not respond to antibiotics. However, their

incubation periods are normally longer than for other biological agents, so incapacitation of

victims may be delayed. Table 2−2 lists the common viral agents along with possible methods

of dissemination, incubation period, symptoms, and treatment.

2.3 Rickettsiae

Rickettsiae are obligate intracellular bacteria that are intermediate in size between most bacteria

and viruses and possess certain characteristics common to both bacteria and viruses. Like

bacteria, they have metabolic enzymes and cell membranes, use oxygen, and are susceptible to

broad-spectrum antibiotics, but like viruses, they grow only in living cells. Most rickettsiae can

be spread only through the bite of infected insects and are not spread through human contact.

Table 2−3 lists the common rickettsiae along with possible methods of dissemination, incubation

periods, symptoms, and treatment.

2.4 Biological Toxins

Biological toxins are poisons produced by living organisms. It is the poison, not the organism,

that produces harmful effects in man. A toxin typically develops naturally in a host organism

(for example, saxitoxin is produced by marine algae); however, genetically altered and/or
6

synthetically manufactured toxins have been produced in a laboratory environment. Biological

toxins are most similar to chemical agents in their dissemination and effectiveness. Table 2−4

lists the common biological toxins along with possible methods of dissemination, incubation

period, symptoms, and treatment.
7

Table 2-1. Bacterial agents

Biological

Agent/Disease Anthrax Brucellosis

E. coli serotype

(O157:H7) Tularemia Cholera

Likely Method

of Dissemination

1. Spores in aerosol

2. Sabotage (food)

1. Aerosol

2. Sabotage

 (food)

Water and food

supply contamination

1. Aerosol

2. Rabbits or ticks

1. Sabotage (food

 and water)

2. Aerosol

Transmissible

Person to

Person

No (except cutaneous) Unknown Unknown, evidence

passed person-toperson in day-care

or nursing homes

No Rare

Incubation

Period

1 d to 43 d 1 wk to 3 wk,

sometimes

months

Unknown 2 d to 10 d 3 d to 5 d

Duration of

Illness

3 d to 5 d (usually

fatal)

Unknown 5 d to 10 d (most

cases)

>2 wk >1 wk

Lethality Contact or cutaneous

anthrax: fatality rate of

5 % to 20 %

Inhalational anthrax:

after symptoms appear

almost always fatal,

regardless of treatment

Low 0 % to 15 % if

develop hemolytic

uremic syndrome

(HUS); 5 % if

develop thrombotic

thrombocytopenic

purpura (TTP)

Moderate if left

untreated

Low (<1 %) with

treatment; high

(>50 %) without

Vaccine

Efficacy

(for aerosol

exposure)/

Antitoxin

Currently no human

data

Vaccine under

evaluation

No vaccine No commercially

available vaccine

No data on aerosol

Symptoms and

Effects

Flu-like, upperrespiratory distress;

fever and shock in 3 d

to 5 d, followed by

death

Irregular

prolonged fever,

profuse sweating,

chills, joint and

muscle pain,

persistent fatigue

Gastrointestinal

(diarrhea,

vomiting)

dehydration; in

severe cases,

cardiac arrest and

death, HUS, or

TTP

Chills; sustained

fever; prostration;

tendency for

pneumonia;

enlarged, painful

lymph nodes;

headache; malaise;

anorexia;

nonproductive

cough

Sudden onset with

nausea, vomiting,

diarrhea, rapid

dehydration,

toxemia and

collapse

Treatment Vaccine available for

cutaneous, possibly

inhalation, anthrax.

Cutaneous anthrax

responds to antibiotics

(penicillin, terramycin,

chloromycetin),

sulfadiazine, and

immune serum.

Pulmonary (inhaled)

anthrax responds to

immune serum in

initial stages but is

little use after disease

is well established.

Intestinal, same as for

pulmonary

Antibiotics Antibiotics

available; most

recover without

antibiotics within

5 d to 10 d; do not

use antidiarrheal

agents

Vaccination using

live attenuated

organisms reduces

severity and

transmittability;

antibiotics

(streptomycin,

aureomycin,

chloromycetin,

doxycycline,

tetracycline, and

chloramphenical)

Replenish fluids and

electrolytes;

antibiotics

(tetracycline,

ciprofloxicin, and

erythromycin)

enhance

effectiveness of

rehydration and

reduce organism in

body

Potential as

Biological

Agent

High, Iraqi and USSR

biological programs

worked to develop

anthrax as a bioweapon

Unknown Unknown High, if delivered

via aerosol form

(highly infectious,

90 % to 100 %)

Not appropriate for

aerosol delivery
8

Table 2-1. Bacterial agents-Continued

Biological

Agent/Disease Diphtheria Glanders Melioidosis

Plague (Bubonic

and Pneumonic) Typhoid Fever

Likely Method

of Dissemination

Unknown 1. Aerosol

2. Cutaneous

1. Food

contamination

(rodent feces)

2.Inhalation

3.Insect bites

4. Direct contact

with infected

animals

1. Infected fleas

(Bubonic and

Pneumonic)

2. Aerosol

(Pneumonic)

1. Contact with

infected person

2. Contact with

contaminated

substances

Transmissible

Person to

Person

High High No High (Pneumonic) High

Incubation

Period

2 d to 5 d 3 d to 5 d Days 1 d to 3 d 7 d to 14 d

Duration of

Illness

Unknown Unknown 4 d to 20 d 1 d to 6 d (usually

fatal)

Unknown

Lethality 5 % to 10 % fatality 50 % to 70 % Variable 5 % to 10 % if

treated

Bubonic: 30 % to

75 % if untreated

Pneumonic: 95 % if

untreated

<1 % if treated;

10 % to 14 % if

untreated

Vaccine

Efficacy

(for aerosol

exposure)/

Antitoxin

DPT vaccine 85 %

effective; booster

recommended every

10 yr

No vaccine No vaccine Vaccine not

available

Oral vaccine (Vivotif)

and single dose

injectable vaccine

(capsular polysaccharide antigen);

both vaccines are

equally effective and

offer 65 % to 75 %

protection against the

disease

Symptoms and

Effects

Local infection usually

in respiratory passages;

delay in treatment can

cause damage to heart,

kidneys, and central

nervous system

Skin lesions,

ulcers in skin,

mucous

membranes, and

viscera; if

inhaled, upper

respiratory tract

involvement

Cough, fever,

chills, muscle/joint

pain, nausea, and

vomiting;

progressing to

death

Enlarged lymph

nodes in groin;

septicemic (spleen,

lungs, meninges

affected)

Prolonged fever,

lymph tissue

involvement;

ulceration of

intestines;

enlargement of

spleen; rose-colored

spots on skin;

constipation or

diarrhea

Treatment Antitoxin extremely

effective; antibiotic

(penicillin) shortens the

duration of illness

Drug therapy

(streptomycin and

sulfadiazine) is

somewhat

effective

Antibiotics

(doxycycline,

chlorothenicol,

tetracycline) and

sulfadiazine

Doxycycline (100

mg 2x/d for 7 d);

ciprofloxicin also

effective

Antibiotics

(amoxicillin or

cotrimoxazole)

shorten period of

communicability and

cure disease rapidly

Potential as

Biological

Agent

Very low––symptoms

not severe enough to

incapacitate; rare cases

of severe infection

Unknown Moderate––rare

disease, no vaccine

available

High––highly

infectious,

particularly in

pneumonic (aerosol)

form; lack of

stability and loss of

virulence

complicate its use

Not likely to be

deployed via aerosol;

more likely for covert

contamination of

water or food
9

Table 2-2. Viral agents

Biological

Agent/Disease Marburg Virus Junin Virus

Rift Valley

Fever Virus Smallpox

Venezuelan Equine

Encephalitis

Likely Method of

Dissemination

Aerosol Epidemiology not

known

Mosquito-borne;

in biological

scenario, aerosols

or droplets

Aerosol 1. Aerosol

2. Infected vectors

Transmissible

Person to Person

Unknown Unknown Unknown High No

Incubation Period 5 d to 7 d 7 d to 16 d 2 d to 5 d 10 d to 12 d 1 d to 6 d

Duration of

Illness

Unknown 16 d 2 d to 5 d 4 wk Days to weeks

Lethality 25 % 18 % <1 % 20 % to 40 %

(Viriole major)

<1 % (Viriole

minor)

1 % to 60 %

Vaccine Efficacy

(for aerosol

exposure)/

Antitoxin

No vaccine No vaccine Inactivated vaccine

available in limited

quantities

Vaccine protects

against infection

within 3 d to 5 d of

exposure

Experimental only:

TC−83 protects

against 30 LD50s to

500 LD50s in

hamsters

Symptoms and

Effects

Sudden onset of

fever, malaise,

muscle pain,

headache, and

conjunctivitis,

followed by sore

throat, vomiting,

diarrhea, rash, and

both internal and

external bleeding

(begins 5th day).

Liver function may

be abnormal and

platelet function

may be impaired.

Hemorrhagic

syndrome, chills,

sweating,

exhaustion and

stupor

Febrile illness,

sometimes

abdominal

tenderness; rarely

shock, ocular

problems

Sudden onset of

fever, headache,

backache, vomiting,

marked prostration,

and delirium; small

blisters form crusts

which fall off 10 d

to 40 d after first

lesions appear;

opportunistic

infection

Sudden illness with

malaise, spiking

fevers, rigors, severe

headache,

photophobia, and

myalgias

Treatment No specific

treatment exists.

Severe cases require

intensive supportive

care, as patients are

frequently

dehydrated and in

need of intravenous

fluids.

No specific

therapy;

supportive

therapy essential

No studies, but IV

ribavirin (30

mg/kg/6 h for 4 d,

then 7.5 mg/kg/8 h

for 6 d) should be

effective

Vaccinia immune

globulin (VIG) and

supportive therapy

Supportive

treatments only

Potential as

Biological Agent

High––actually

weaponized by

former Soviet

Union biological

program

Unknown Difficulties with

mosquitos as

vectors

Possible, especially

since routine

smallpox

vaccination

programs have been

eliminated worldwide (part of USSR

offense bioprogram)

High––former U.S.

and U.S.S.R.

offensive biological

programs

weaponized both

liquid and dry forms

for aerosol

distribution
10

Table 2-2. Viral agents-Continued

Biological

Agent/Disease Yellow Fever Virus Dengue Fever Virus Ebola Virus

Congo-Crimean

Hemorrhagic

 Fever Virus

Likely Method of

Dissemination

Mosquito-borne Mosquito-borne 1. Direct contact

2. Aerosol (BA)

Unknown

Transmissible

Person to Person

No No Moderate Yes

Incubation Period 3 d to 6 d 3 d to 15 d 4 d to 16 d 7 d to 12 d

Duration of

Illness

2 wk 1 wk Death between 7 d to

16 d

9 d to 12 d

Lethality 10 % to 20 % death in

severe cases or full

recovery after 2 d to

3 d

5 % average case

fatality by producing

shock and hemorrhage,

leading to death

High for Zaire strain;

moderate with Sudan

15 % to 20 %

Vaccine Efficacy

(for aerosol

exposure)/

Antitoxin

Vaccine available;

confers immunity for

>10 yr

Vaccine available No vaccine No vaccine available;

prophylactic ribavirin

may be effective

Symptoms and

Effects

Sudden onset of chills,

fever, prostration,

aches, muscular pain,

congestion, severe

gastrointestinal

disturbances, liver

damage and jaundice;

hemorrhage from skin

and gums

Sudden onset of fever,

chills, intense

headache, pain behind

eyes, joint and muscle

pain, exhaustion and

prostration

Mild febrile illness,

then vomiting,

diarrhea, rash, kidney

and liver failure,

internal and external

hemorrhage (begins 5th

day), and petechiae

Fever, easy bleeding,

petechiae, hypotension

and shock; flushing of

face and chest, edema,

vomiting, diarrhea

Treatment No specific treatment;

supportive treatment

(bed rest and fluids)

for even the mildest

cases

No specific therapy;

supportive therapy

essential

No specific therapy;

supportive therapy

essential

No specific treatment

Potential as

Biological Agent

High, if efficient

dissemination device is

employed

Unknown Former Soviet Union Unknown
11

Table 2-3. Rickettsiae

Biological

Agent/Disease Endemic Typhus Epidemic Typhus Q Fever

Rocky Mountain

Spotted Fever

Likely Method of

Dissemination

1. Contaminated

feces

2. Infected insect

larvae

3. Rat or flea bites

1. Contaminated feces

2. Infected insect larvae

1. Sabotage (food

supply)

2. Aerosol

Infected wood ticks

Transmissible

Person to Person

No No Rare No

Incubation

 Period

6 d to 14 d 6 d to 15 d 14 d to 26 d 3 d to 14 d

Duration

of Illness

Unknown Unknown Weeks Unknown

Lethality 1 %, increasing in

people >50 yr old

10 % to 40 %

untreated; increases

with age

Very low 15 % to 20 % untreated

(higher in adults);

treated—death rare

with specific therapy

(tetracycline or

chloramphenicol)

Vaccine Efficacy

(for aerosol

exposure)/

Antitoxin

Unknown Vaccine confers

protection of uncertain

duration

94 % protection

against 3500 LD50s in

guinea pigs

No vaccine

Symptoms

and Effects

Sudden onset of

headache, chills,

prostration, fever,

pain; maculae

eruption on 5th day to

6

th day on upper

body, spreading to all

but palms, soles, or

face, but milder than

epidemic form

Sudden onset of

headache, chills,

prostration, fever, pain;

maculae eruption on 5th

day to 6th day on upper

body, spreading to all

but palms, soles, or

face

Mild symptoms

(chills, headaches,

fever, chest pains,

perspiration, loss of

appetite)

Fever and joint pain,

muscular pain; skin

rash that spreads

rapidly from ankles and

wrists to legs, arms,

and chest; aversion to

light

Treatment Antibiotics

(tetracycline and

chloramphenicol);

supportive treatment

and prevention of

secondary infections

Antibiotics

(tetracycline and

chloramphenicol);

supportive treatment

and prevention of

secondary infections

Tetracycline (500 mg/

6 h, 5 d to 7 d) or

doxycycline (100 mg/

12 h, 5 d to 7 d) also,

combined

Erthyromycin

(500 mg/6 h) and

rifampin (600 mg/d)

Antibiotics—

tetracycline or

chloramphenicol

Potential as

Biological Agent

Uncertain––broad

range of incubation

(6 d to 14 d) period

could cause infection

of force deploying

biological agent

Uncertain––broad

range of incubation

(6 d to 14 d) period

could cause infection of

force deploying

biological agent

Highly infectious, is

delivered in aerosol

form. Dried agent is

very stable; stable in

aerosol form.

Unknown
12

Table 2-4. Biological toxins

Biological

Agent/Disease Botulinum Toxin

Staphylococcal

enterotoxin B

Tricothecene

mycotoxins

Ricin (Isolated

from Castor Beans) Saxitoxin

Likely

Method of

Dissemination

1. Aerosol

2. Sabotage (food

and water)

1. Sabotage (food

supply)

2. Aerosol

1. Aerosol

2. Sabotage

1. Aerosol

2. Sabotage (food &

water)

Contaminated

shellfish; in

biological scenario,

inhalation or toxic

projectile

Transmissible

Person to Person

No No No No No

Incubation

 Period

Variable (hours to

days)

3 h to 12 h 2 h to 4 h Hours to days 5 min to 1 h

Duration

of Illness

Death in 24 h to

72 h; lasts months

if not lethal

Hours Days to months Days––death within

10 d to 12 d for

ingestion

Death in 2 h to 12 h

Lethality 5 % to 60 %,

untreated

<5 % treated

<1 % Moderate 100 %, without

treatment

High without

respiratory support

Vaccine Efficacy

(for aerosol

exposure)/

Antitoxin

Botulism antitoxin

(IND)

Prophylaxis toxoid

(IND)

Toxolide

No vaccine No vaccine No vaccine No vaccine

Symptoms

and Effects

Ptosis; weakness,

dizziness, dry

mouth and throat,

blurred vision and

diplopia, flaccid

paralysis

Sudden chills,

fever, headache,

myalgia,

nonproductive

cough, nausea,

vomiting and

diarrhea

Skin––pain,

pruritis, redness

and vesicles,

sloughing of

epidermis;

respiratory––nose

and throat pain,

discharge,

sneezing,

coughing, chest

pain, hemoptysis

Weakness, fever,

cough, pulmonary

edema, severe

respiratory distress

Light headedness,

tingling of

extremities, visual

disturbances,

memory loss,

respiratory distress,

death

Treatment Antitoxin with

respiratory support

(ventilation)

Pain relievers and

cough suppressants

for mild cases; for

severe cases, may

need mechanical

breathing and fluid

replenishment

No specific

antidote or

therapeutic

regimen is

available;

supportive and

symptomatic care

Oxygen, plus drugs

to reduce

inflammation and

support cardiac and

circulatory

functions; if

ingested, empty the

stomach and

intestines; replace

lost fluids

Induce vomiting,

provide respiratory

care, including

artificial respiration

Potential

as Biological

Agent

Not very toxic via

aerosol route;

extremely lethal if

delivered orally.

Since covert

poisoning is

indistinguishable

from natural

botulism,

poisoning could

have limited use

Moderate––could

be used in food

and limited

amounts of water

(for example, at

salad bars); LD50 is

sufficiently small

to prevent

detection

High––used in

aerosol form

(“yellow rain”) in

Laos, Kampuchea

and Afghanistan

(through 1981)

Has been used in

1978––Markov

murder (see app. A,

ref. 6); included on

prohibited Schedule

I chemicals list for

Chemical Weapons

Convention; high

potential for use in

aerosol form

Moderate, aerosol

form is highly toxic
13

3. CHALLENGES TO BIOLOGICAL AGENT DETECTION

Biological agents are effective in very low doses. Therefore, biological agent detection systems

need to exhibit high sensitivity (i.e., be able to detect very small amounts of biological agents).

The complex and rapidly changing environmental background also requires these detection

systems to exhibit a high degree of selectivity (i.e., be able to discriminate biological agents

from other harmless biological and nonbiological material present in the environment). A third

challenge that needs to be addressed is speed or response. These combined requirements

provide a significant technical challenge. Additionally, there has been limited development in

the area of biological agent detection equipment in the commercial market (i.e., hand-held

devices). There are several detection systems being developed and tested by the military that

show promise. However, these systems are relatively complicated, require training for

successful operation and maintenance, and are expensive to purchase and operate. It is expected

that over the course of the next 5 years, commercial instrumentation, hardened for use in the

field, may become available at reasonable costs.

The purpose of this section is to identify some of the major challenges associated with biological

agent detection. Specifically, section 3.1 addresses challenges associated with the ambient

environment, section 3.2 discusses challenges with selectivity, section 3.3 discusses challenges

with sensitivity, and section 3.4 addresses challenges with sampling.

3.1 The Ambient Environment

The environment in which we live and operate is an extremely complex and dynamic medium.

The meteorological, physical, chemical, and biological constituents of a “normal” atmospheric

environment all impact our ability to detect biological agents. In order to understand the

complex effect that the ambient environment can have on biological agent detection, the

remainder of this section discusses specifics of the particulate background, the biological

background, and the optical background, respectively.

3.1.1 The Particulate Background

Particulates in the atmosphere originate from a number of sources. Dust, dirt, pollen, and fog are

all examples of naturally occurring particulates found in the air. Man-made particulates such as

engine exhaust, smoke, and industrial effluents (smokestacks) also contribute significantly to the

environmental particulate background. Therefore, the particulate background can be defined as

the combination of natural and man-made particles in the atmosphere that are nonpathogenic

(does not cause disease) in nature. Biological agents (not including toxins) consist of

particulates of pathogenic (disease causing) cells. The particulate background can change on a

minute-by-minute basis depending on the meteorological conditions at the time. For example,

the particulate background next to a road will change dramatically depending on whether there is

traffic on the road disturbing the dust, or if the road is empty. Likewise, if there is little wind,

not many particulates are carried into the atmosphere; however, when the wind begins to blow, it

can carry many particulates from the immediate vicinity, as well as from remote locations. The

challenge for a biological detection system is to be able to discriminate between all of the

naturally occurring particulates and the biological agent particulates.
14

Particle counters can be used to monitor changes in the particulate background on a real-time

basis because these systems see particles in the air and can count them. If the number of

particles increases rapidly, it is possible that biological agents are being used; however, it must

be stressed that particle counters cannot determine if the particulates are dust, pollen,

engine exhaust, or biological agents. Other, more sensitive and selective, tests must be

performed on the particulates to determine if biological agents are present. Particle counters are

best used in a detection system where the particle counter activates a sampler that collects a

sample of the particles for a more detailed analysis.

3.1.2 The Biological Background

Our environment is filled with living creatures that form a large and complex biological

background from which we must identify biological agents. The challenge for a biological agent

detection system is to be able to pick out a specific signal from the biological agent while

rejecting, or at best minimizing, any signals originating from the nonpathogenic (nontoxic)

biological background. This is a significant challenge given the amount of biological

particulates in the environment. Research has identified a variety of potential bio-aerosol

sources (i.e., adjoining crop fields that are fertilized with “night soil,” garbage incinerators,

landfills, industrial areas, and dairy farms). Studies have shown that the concentration of bioaerosols depends on the location of the measurement. In Oregon, a study showed that the

concentration of bio-aerosol in an urban setting was six times greater than along the coast and

almost three times greater than in a rural setting.

Data shown in figure 3−1 suggest that not only do biological aerosols vary by location, they also

vary significantly by time of day.

Figure 3-1. Airborne bacterial concentration fluctuation in a single day4

 

4Aerosolized bacterial concentration fluctuation over a 24 h period. The vertical (y) axis is bacterial concentration per cubic meter of air. The

horizontal (x) axis is the time of day; shaded regions represent nighttime hours, and the clear region is daytime hours. The graph shows that in

the early morning hours, the airborne bacterial concentration is low, but it increases rapidly during daylight, reaching a maximum at 8:00 a.m. It

then falls to a lower level for most of the day and significantly increases towards the end of the day.
15

3.1.3 The Optical Background

Systems such as laser or passive infrared (IR) systems rely on optical properties for detection of

biological agents. They can be affected by micron range particulates, as well as by other

obstructions to visibility such as rain, fog, snow, and dust. Aerosols and precipitation may act

like mirrors, reflecting and diffusing the light energy to and from the detector, and in the case of

some aerosols, return false signatures (e.g., fluorescence from engine exhaust and pollens may

confuse some ultraviolet (UV) based systems). Consequently, different standoff systems are

affected to different degrees by precipitation and aerosols. Infrared-based systems, as a rule,

tend to be less affected by atmospheric clarity than UV-based systems.

3.2 Selectivity of the Detection System

Detection systems must exhibit a high degree of selectivity for biological agents. The selectivity

of a detection system can be defined as its ability to discriminate between the target agent and

the environmental interferants. The degree to which the selectivity of a system is affected by

interferants depends on the type of measurement being conducted. For example, dust and pollen

can be considered interferants for a particle counter, while water vapor and fog are interferants

for standoff IR detection systems. For biological agent monitoring, the most difficult interferants

originate from the biological background (i.e., live nonpathogenic matter). Generally, the more

selective systems require more sample processing and multiple detectors. A single system for

detection of biological agents in the environment that exhibits high selectivity currently does not

exist as a commercially available item. The selective systems currently developed by the

military are limited to detection of a small number of agents and are prohibitively expensive.

3.3 Sensitivity

Detection systems must exhibit high sensitivity for the biological agents because of the agent’s

low effective doses (fig. 1). Sensitivity can be defined as the smallest amount of target agent that

gives a reproducible response above the system noise for a detector. The system noise can be

defined as the random fluctuation of the detector response and is generally associated with small

variations in electronic output. Other noise that degrades the sensitivity is caused by interferants

in the environment. In a perfect detection system, the system sensitivity (only dependent on the

electronic noise) defines how much of the target agent can be detected. Interferants cause the

sensitivity to decrease because the system needs more of the target agent to distinguish it from

the interferants.

3.4 Sampling

The primary infection route from exposure to biological agents is through inhalation, and it is

likely that most of the initial aerosol would have settled by the time emergency first responders

arrive on the scene of an incident. This does not lessen the possibility of infection of the first

responders by reaersolization of the agent but requires that the emergency first responders take

more than just air samples for analysis. It may be critical for the emergency first responders to

conduct environmental (soil/water) sampling and air and swipe tests to corroborate the

occurrence of a biological attack and to determine if the biological agent is still present.
16

Emergency first responders may only be involved in post-incident activities and may not have

any need for early warning capabilities.

Since sampling is a key issue for all analytical devices, the way a sample is taken and how it is

handled will affect the outcome of the analysis. In a point collection/detection scenario,

sampling for biological agent particlates in the air is especially difficult due to the low effective

doses of these agents. To sample biological agents effectively, samplers are used that pass large

volumes of air through the sampler, dispersing the small amount of agent contained in a large

volume of air into a small volume of water, thereby forming a concentrated mixture of

particulates in water. By concentrating the biological particulates, current detection systems that

are not able to detect biological agents at low dose levels can detect the biological agents in the

concentrated mixture.
17

4. BIOLOGICAL DETECTION SYSTEM COMPONENTS

The effective detection of biological agents in the environment requires a multicomponent

analysis system because of the complexity of the environment. Other variables contributing to

the effectiveness of detection of biological agents are the detection process itself and the efficient

use of consumables in the field. Biological agent detection systems generally consist of four

components: the trigger/cue, the collector, the detector, and the identifier. Figure 4−1 shows a

flow diagram for a typical point detection automated architecture system. The function of these

components is described in the remainder of this section, while section 5 will provide

representative examples of each component.

Figure 4–1. Typical point detection automated architecture

(with a combined trigger/cue)

4.1 Trigger/Cue

Trigger technology is the first level of detection that determines any change in the particulate

background at the sensor, indicating a possible introduction of biological agents. Detection of an

increase in the particulate concentration by the trigger causes the remaining components of the

detection system to begin operation. The trigger function typically provides a means of

continuously monitoring the air without unnecessary use of consumables, thus keeping the

logistical burden of biological agent detection low.

To reduce false positives (alarm with no biological agent) and false negatives (no alarm with

agent), many detection systems combine trigger technology with a second detector technology

(such as fluorescence that provides more selectivity) into a single technology known as cueing.

Most effective cueing technologies can detect airborne particulates in near real time and can

discriminate between biological agent aerosol particles and other particles in air, avoiding

Tiered detection approach to reduce consumable usage

Quick, generic detection followed by longer, specific identification

Typical Point Detection

Automated Architecture

 Bio/

No Bio

Is It Safe?

Fluid

Initiate
18

unnecessary system activation. For example, a cueing device monitors the air for particulates as

does any other trigger device. When the particulate concentration increases, the cue determines

if the particulates are biological in nature. The cue device generally uses a fluorescence detector

to make this determination. If the particulates are found to be biological, the cue device activates

the collector for sample collection.

4.2 Collector

As discussed in section 3.4, sampling of the biological agent is a crucial part of the identification

system. The effective dose for some agents is extremely small; therefore, highly efficient

collection devices must be employed. One type of collector pumps large volumes of air through

a chamber where the air mixes with water. The water scrubs all the particulates from the air,

resulting in a sample containing particulates suspended in water. Once collected in the water, the

sample is further concentrated by evaporation of a portion of the water. After concentration, the

sample moves into the analytical section of the biological agent detection system.

4.3 Detector

Once a sample has been collected/concentrated, it must be determined if the particulates are

biological or inorganic in origin. To accomplish this, the sample is passed to a generic detection

component that analyzes the aerosol particles to determine if they are biological in origin. This

component may also classify the suspect aerosol by broad category (e.g., spore, bacterium,

toxin/macromolecule, or virus). In its simplest form, the detector acts as a “gateway” for further

analysis. If the sample exhibits characteristics of biological particles, it is passed through to the

next level of analysis. If the sample does not exhibit such characteristics, it is not passed to the

next level of analysis, thereby conserving analytical consumables.

It is important to note that detection has traditionally taken place after the trigger function. For

example, an aerosol particle sizer (APS) triggers, then a detector (e.g., flow cytometer) examines

the aerosol for biological content. Many of the newer detection technologies combine the trigger

and detection functionalities into a single instrument, creating a cueing instrument. As described

in section 4.1, the cue first detects a rise in particulates then determines if the particulates are of

biological origin. If the sample is biological, the collector gathers a sample and passes it directly

to the identifier.

4.4 Identifier

An identifier is a device that specifically identifies the type of biological agent collected by the

system. Identifiers are generally limited to a preselected set of agents and cannot identify agents

outside of this set without the addition of new identifier chemistry/equipment or

preprogramming. Because the identifier performs the final and highest level of agent detection,

it is the most critical component of the detection architecture and has the widest variety of

technologies and equipment available. The information obtained from the identifier is then used

to determine protection requirements and treatment of exposed personnel.
19

5. OVERVIEW OF BIOLOGICAL AGENT DETECTION

SYSTEM TECHNOLOGIES

The applicability of biological agent detection equipment to emergency first responders will

depend on the characteristics of the detection equipment, the type of biological agent to be

detected, and the objective of the emergency first responder unit. Good analytical results from

the various analyzers will depend on the ability to effectively sample the environment and

deliver the biological agent to the analyzer.

Biological detection systems are currently in the research and early development stages.

There are some commercially available devices that have limited utility (responding only

to a small number of agents) and are generally high cost items. Because commercially

available biological warfare (BW) detection systems and/or components exhibit limited

utility in detecting and identifying BW agents and are also costly, it is strongly

recommended that first responders be very careful when considering a purchase of any

device that claims to detect BW agents. This is a very different situation when compared to

chemical detection equipment; there are various technologies for detection of chemical agents

and toxic industrial materials (TIMs) that can be purchased by the emergency first responder.

One reason for the lack of available biological detection equipment is that detection of

biological agents requires extremely high sensitivity (because of the very low effective dose

needed to cause infection and spread the disease) and an unusually high degree of selectivity

(because of the large and diverse biological background in the environment).

Another reason for the lack of biological detection equipment is that biological agents,

compared to chemical agents, are very complex systems of molecules, which makes them much

more difficult to identify. For example, Ionization/Ion Mobility Spectrometry (IMS), an

excellent (though expensive) system for collection, detection, and identification of chemical

agents, cannot detect or discriminate biological agents in its present form. In fact, the need for

high-efficiency collection and concentration of the sample, high sensitivities, and high

selectivities make all chemical detectors in their current form unusable for biological agent

detection.

Because of the need for high selectivity and sensitivity, the biological detection systems are

necessarily complex devices consisting of various subunits. Each subunit performs a specific

collection, detection, and identification task. In this section, the various units and subunits that

make up biological agent point and standoff detection systems are described. Specifically,

section 5.1 discusses the separate technologies utilized with point detection, section 5.2

discusses standoff technologies (both short range and long range), and section 5.3 addresses

passive standoff detection.

For reference only, examples of the size and complexity of integrated biological detection systems

are presented in figure 5−1 and figure 5−2. They are the Biological Integrated Detection Systems

(BIDS) from the United States and a cutaway picture of the Integrated Biological Integrated

Detection System from the United Kingdom, respectively.
20

Figure 5-1. Biological Integrated

Detection System (BIDS)

Figure 5-2. Cutaway of the UK Integrated

Biological Detection System (IBDS)

5.1 Point Detection Technologies

Point detectors are those sensors that must be in the aerosol plume or have the suspect biological

agent introduced into/onto them for sensing. Point detection systems have traditionally

encompassed the following components: trigger/cue (nonspecific biological agent detectors),

sampler/collector, and identifier (specific identification technologies).

5.1.1 Trigger/Cue (Nonspecific Biological Agent Detectors)

The function of the trigger is to provide early warning that a change in the background air has

occurred. Operation of a trigger requires establishing background aerosol levels in a specific

location and then sensing that an increase in the aerosol particle count in the background has

occurred. A trigger is nonselective and does not identify the organism but only indicates a

change in the background aerosol level. Since a trigger is nonselective, a detector is required if

there is no cue.

A cueing device is first able to determine when there is an increase in particulates and then is

able to distinguish between concentrations of biological aerosols and nonbiological aerosols

(nonspecific biological agent detection). Descriptions of several detector technologies are

presented in section 5.1.3.

Brief descriptions of trigger/cue technologies are presented in the section below.

5.1.1.1 Particle Measurement

One technique used for nonspecific detection is counting the relative number of particles in

specific size ranges (typically 0.5 μm to 30 μm). A variety of technologies are used for particle

monitoring and/or counting, but aerodynamic particle sizing has been directly applied to field

biological agent detection. Several examples of particle measurement technologies follow.

Aerodynamic Particle Sizing (APS): The particle-laden air stream is drawn into the APS device

through a flow nozzle, producing a controlled high-speed aerosol jet. During the measurement
21

period, the air velocity remains constant but because of the different sizes of the individual

particles within the jet, they accelerate at different rates based on their relative sizes (smaller

particles accelerate faster than larger particles). A laser beam measures the time of flight of the

individual particles.

High Volume Aerodynamic Particle Sizer (HVAPS): The HVAPS passes an accelerated,

concentrated air stream past a laser-based particle counter to obtain aerosol particle size

distribution and concentration. This instrument cannot discriminate biological from nonbiological aerosols.

Met-One: The Met-One is a compact, low-power aerosol particle sizer and counter about the

size of a large, hand-held calculator. This device is available commercially and is typically used

to monitor clean rooms. The Met-One draws an air sample through a laser-illuminated sample

volume where airborne particles scatter light. The light scattered by individual particles is then

detected using a photodiode. Like the HVAPS, the Met-One looks for a statistically significant

rise in aerosol concentration over background; however, the Met-One is not able to resolve the

particle sizes as finely as the HVAPS. The Met-One gains its size and weight savings through a

combination of low airflow and use of a low-power, diode laser.

5.1.1.2 Fluorescence Methods

Fluorescence approaches involve excitation of molecular components of a material with light,

usually in the ultra violet (UV) region of the spectrum. The excited component spontaneously

reverts to an unexcited state followed by emission of light at different wavelengths. Because the

emission spectrum is specific to the molecular component being irradiated and the excitation

wavelength, this phenomenon can be exploited in detection of biological material

(biofluorescence). Biofluorescence-based techniques generate data from only some specific

molecular components of biological material, allowing it to be a tool for nonspecific agent

detection by providing the emission spectrum of a common material (i.e., tryptophan) when an

unknown sample is irradiated.

The two types of fluorescence measurement approaches are primary and secondary. In primary

biofluorescence, some common, naturally fluorescent component of biomaterials, such as

tryptophan (an amino acid building block of protein), is measured. Secondary fluorescence

methods involve introducing (tagging) a special fluorophore (i.e., fluorochrome stain) to the

sample before UV irradiation. Secondary methods require a longer measurement time and add

complexity to the measurement process. Several devices that use biofluorescence technologies

are included in the remainder of this section.

Fluorescent Aerodynamic Particle Sizer (FLAPS): FLAPS is an Aerodynamic Particle Sizer

(APS) that has been modified to include an additional laser (blue or UV wavelength) that

provides for aerosol particle fluorescence in addition to standard particle size information.

Besides obtaining the aerodynamic particle size, the laser’s signal acts as a trigger to open a time

window in which to look for particle fluorescence. The information obtained from this

technology will be more specific than the current standard particle size and number density

results.
22

The FLAPS II device is part of the Canadian Integrated Biological Agent Detection System

(CIBADS), a.k.a., the 4WARN detection suite. The CIBADS is an integrated system of

components developed by the Canadian Ministry of Defense that currently contains a

detector/trigger function, sample collection function, meteorological instrumentation, and

communications equipment. A picture of the FLAPS II is presented in figure 5−3, and the

4WARN system from Canada is presented in figure 5−4.

Figure 5-3. FLAPS II (component

of the Canadian 4WARN System)

Figure 5-4. Canadian Integrated

Biological-Chemical Agent Detection

System (CIBADS)/4WARN

A variation of the FLAPS particle sizer is the Ultra Violet Aerodynamic Particle Sizer (UVAPS) that

uses time-of-flight particle sizing, light scattering, and UV fluorescence intensity to nonspecifically

detect biological agents in air samples. The UVAPS (as well as the FLAPS) is commercially available

from TSI Inc., Particle Instruments.

The Biological Aerosol Warning System (BAWS) is effective as a trigger/cue technology. The

BAWS uses a micro-laser-based system that analyzes two biological fluorescence wavelengths

to determine if an unusual biological event is happening. The BAWS does not count aerosol

particles. It can detect in real time and can discriminate biological agent aerosol particles from

other particles in the air to avoid false triggers.

A technique called Portable Biofluorosensor (PBS) was used during Operation Desert Storm.

The technique used UV light from a xenon flash lamp to excite airborne aerosols and aerosols

dissolved in water. The excitation wavelength minimized interference from dust, exhaust, etc.,

but did not eliminate false positives. Liquid samples containing spores provided better analysis

results than airborne samples.

The Single-Particle Fluorescence Counter (SPFC), developed by the Naval Research Laboratory

(NRL), employs continuous airflow across a 780 nm laser-diode beam, resulting in light

scattering from individual aerosol particles in the air. The total intensity of scattered light is
23

measured, and particle size is calculated. This event also triggers a 266 nm UV laser pulse that

causes fluorescent particles to emit light at a different wavelength (i.e., the particles fluoresce).

5.1.2 Samplers/Collectors

Since an extremely low airborne concentration of biological agents can be difficult to detect but

still cause severe effects, a device to concentrate particles/aerosols in the air stream is needed.

A collector/concentrator samples the atmosphere and concentrates the airborne particles into a

liquid medium for analysis. Several types of samplers/collectors have been evaluated for

biological agent detection. The principal differences between collection for biological agent

detection and other types of aerosol or particulate sampling are (1) biological agent sampling is

normally targeted at living organisms, so the sampling techniques must preserve and not harm

the collected sample; (2) most biological detection and identification technologies require a

liquid sample, so the collection must be from an aerosol or particulate in a liquid; and (3) the

liquid sample must be highly concentrated and available for rapid analysis since response time

is critical.

A collector is most useful when it is part of a detection system. When the collector receives a

signal from a trigger indicating a change in the background level, an air sample is collected, and

airborne particles are concentrated into a liquid medium.

The efficiency of a collector at capturing and concentrating aerosol samples typically affects

several downstream functions. In virtually all systems, the collectors feed into the identification

component of the biological detection system and also provide the samples that are used for

confirmatory identification and forensic analysis.

Collectors can be broadly divided into two groups. One group contains collectors that are large

and consume much power. These collectors, on the whole, have a high collection and

concentration efficiency and are candidates for detection systems that operate well away from

the line or point of agent release. The other group contains those collectors that consume little

power, are hand-portable, and have relatively low collection and concentration efficiencies.

Whereas these collectors would work well in high agent concentrations (e.g., near the point or

line of release, or perhaps indoors), they would fail to provide an adequate sample to

downstream instruments. It should also be noted that collectors significantly contribute to the

overall weight, size, and power requirements of a detection system.

Examples of sampler/collector technologies include Viable Particle Size Samplers (Impactors),

Virtual Impactors, Cyclones, and Bubblers/Impingers.

5.1.2.1 Viable Particle Size Samplers (Impactors)

A conventional impactor operates by accelerating an air stream of particles through a nozzle and

diverting the air stream against an impaction plate maintained at a fixed distance from the nozzle.

The larger particles are unable to follow the fluid streamlines (air in this case) because of their

large inertia; smaller particles follow the fluid streamlines and exit the sampler.
24

The impactor usually has multiple stages and each stage contains a number of precision-drilled

orifices that are a constant size for each stage. Particle laden air enters the instrument, and the

airborne particles are directed towards the collection surfaces by the jet orifices. Any particle not

collected by a specific stage follows the stream of air around the edge of the collection surface to

the next stage. The collection plate is typically a petri dish with selective agar (selective to a

specific organism). The plates are incubated (typically 24 h to 48 h) and after incubation, the

number of colonies on each plate are counted.

5.1.2.2 Virtual Impactors

A virtual impactor is similar to a conventional impactor but uses a different impaction surface.

The flat plate of the conventional impactor is replaced by a collection probe, and the larger

particles penetrate the collection probe instead of striking a flat plate. By properly controlling

the airflow in the impactor, it is possible to collect particles in a specific size range. In addition,

the final stage can then aim the particle stream onto a liquid, resulting in a highly concentrated

liquid sample.

The Liquid Sampler (PEM-0020) with carousel is manufactured by Power Engineering and

Manufacturing, Inc. The device uses virtual impaction to collect and concentrate airborne

particles onto liquid film. The operator can select the number of samples to be collected (up to

10) and can choose from several preprogrammed sampling protocols that vary the volume and

the collection time for each tube. Initiation of the sample collection is by external trigger or

manual push button. The unit automatically repositions the carousel at the end of the collection

cycle. The entire carousel can be quickly removed and replaced.

The BioVICTM Aerosol Collector, developed by MesoSystems Technology, Inc., serves as a

front-end air sampler for biological detection systems. It is an impacter that preconcentrates the

air stream, capturing large numbers of particles either into a small volume of liquid, into a small

air stream, or onto a solid surface for delivery into the sensor. The BioVICTM can be used with

PCR, fluorescent-based optical sensors, mass spectrometry, pyrolysis GC mass spectrometry, or

flow cytometry. Figure 5−5 shows a picture of the BioVICTM Aerosol Collector.

Figure 5-5. BioVICTM Aerosol Collector,

MesoSystems Technology, Inc.
25

5.1.2.3 Cyclone Samplers

A cyclone is an inertial device that is commonly used in industrial applications for removing

particles from large airflows. A particle-laden air stream enters the cyclone body and forms an

outer spiral moving downward towards the bottom of the cyclone. The larger particles are

collected on the outer wall due to centrifugal force, and the smaller particles follow the airstream

that forms the inner spiral and leave through the exit tube. Water spray applied to the outer walls

of a cyclone facilitate particle collection and preservation. Several examples of cyclone samplers

are discussed in the remainder of this section.

The Interim Biological Agent Detector System (IBADS) was initially developed for the Navy. It

uses a wetted-wall cyclone to collect the aerosol particles into an aqueous sample. Variants of

this device are in use in the Portal Shield Biological Detection System and in the current version

of the Joint Biological Point Detection System (JBPDS). See figure 5−6 for an example of the

JBPDS.

The Smart Air Sampler System (SASS 2000) is a device that has been independently developed

by Research International and also uses wetted-wall cyclone technology. This hand-held device

can operate on battery power. An example of the SASS 2000 is shown in Figure 5−7.

Figure 5-6. Joint Biological Point

Detection System (JBPDS)

Figure 5-7. Smart Air Sampler System

(SASS 2000), Research International

The Portable High-Throughput Liquid Aerosol Air Sampler System (PHTLAAS) is a small

hand-held device that uses technology similar to the wetted-wall cyclone technology. This

instrument concentrates the contaminants found in a large volume of air into a small volume of

liquid for ultrasensitive semiquantitative detection. Zaromb Research Corporation has

independently developed this device.
26

5.1.2.4 Hand-Held Sampling Kit

The Department of Defense Biological Sampling Kit (DoD BSK) is a prepackaged kit containing

a panel of eight hand-held immunochromatographic assay (HHA) devices (i.e., able to

simultaneously identify up to eight different biological agents), a dropper bottle of buffer

solution, two sterile cotton-tipped swabs, and an instruction card. The DoD BSK is included in

the sampler/collection section because it is used for field screening where the concentration of

agent is expected to be high and not for positive identification. The kit is not to be used for

screening soil samples since some soil constituents can cross-react with the HHA reagents if

present in high enough concentrations. In addition, the DoD BSK should not be used for

screening heavily dust-laden surfaces. Also, the kit is not sensitive enough to detect the minute

amounts of precipitate that may fall out from an attack that originated from a distant location

(e.g., a long line source release from several kilometers away).

The advantages of the DoD BSK are that it is inexpensive, reliable, easy to use, and the assays in

the kit are improved concurrent with the assays in the other detection programs. Disadvantages

of the DoD BSK are that it does not possess a generic detection capability (it is an identifier),

and each kit is for one time use only.

5.1.2.5 Hand-Held Sampling Device

The BioCaptureTM BT-500 Air Sampler was developed by MesoSystems Technology, Inc., and

incorporates the BioVICTM Aerosol Collector, also developed by MesoSystems Technology, Inc.

It is a hand-held, battery-powered air sampler that collects airborne samples for quantifying

concentration levels. The microbes are captured and concentrated into an aqueous sample for

analysis by whole cell rapid detection, nucleic acid, or other liquid-based sensor systems. The

removable single-use cartridge can also be archived for evidence of a biological incident. An

example of the BioCaptureTM BT-500 Air Sampler is shown in figure 5−8.

Figure 5-8. BioCaptureTM BT-500 Air

Sampler, MesoSystems Technology, Inc.
27

5.1.3 Detectors

Detectors are those components/instruments used to determine if the particulates are biological

or inorganic in origin and if further analysis of the sample is needed. Some detectors require

additional processing of a sample before it can be introduced into the detector, while others can

use a sample directly from the environment. In this section, detectors are broadly divided into

two groups, wet detection (flow cytometry) and dry detection (mass spectrometry).

5.1.3.1 Wet Detection (Flow Cytometry)

Cytometry is the measurement of both physical and chemical characteristics of cells. Flow

cytometry (widely used as a wet detector for biological agents) uses the same technique as

cytometry but makes the measurements of cells or other particles present in a moving fluid

stream as they pass through a testing point. It measures particle sizes and counts particles in

liquid suspensions through the use of laser light scattering. Flow cytometers involve

sophisticated fluidics, laser optics, electronic detectors, analog to digital converters, and

computersto provide an automated method for bio-chemical analysis and to process thousands of

cells in a few seconds. Typically, the sample will also be treated by addition of a fluorescent dye

that reacts with biological material (e.g., DNA). Flow cytometers have been commercially

available since the early 1970s and increasingly have been used since then. Examples utilizing

this technology are the Los Alamos National Laboratory Flow Cytometer (LANL) and the

Becton Dickenson Flow Cytometer (FACSCaliber). They will be briefly discussed below.

The Los Alamos National Laboratory (LANL) Flow Cytometer employs a green (HeNe) laser

diode. Particle size is measured by two light-scatter detectors, and fluorescence is measured by

two photomultiplier tubes. This instrument is also known as the “Mini-Flow Cytometer” and is

just 1.15 ft3 in size, 30 lb in weight, and requires 1 kW of power.

The B-D Flow Cytometer FACSCount, manufactured by Becton Dickenson, employs a direct

two-color immunogluorescence method and uses a green (HeNe) laser.

The B-D Flow Cytometer FACSCaliber, manufactured by Becton Dickenson, is a four-color

Modular Analytical Flow Cytometer that uses a 15 mW air-cooled blue argon-ion laser and a red

laser diode. The FACSCalibur also has an optional sorter. Figure 5−9 shows an example of the

B-D Flow Cytometer FACSCaliber.
28

Figure 5-9. B-D Flow Cytometer FACSCaliber,

Becton Dickenson

5.1.3.2 Dry Detectors (Mass Spectrometry)

Mass spectrometry (MS) is a microanalytical technique that requires only a few nanograms of

analyte to obtain characteristic information on the structure and molecular weight of the analyte.

The technique ionizes molecules and breaks them apart into characteristic fragments (the

fragmentation pattern constitutes its “mass spectrum”). The mass spectrometer requires that

samples be introduced in the gaseous state. Sample introduction into the mass spectrometer can

be by direct air/gas sampling, a direct insertion probe, membrane inlets, effluent from a gas

chromatograph (GC), effluent from a high-performance liquid chromatograph (HPLC), capillary

electrophoresis, and effluent from pyrolysis devices. Several examples of detection equipment

utilizing mass spectrometry are discussed below.

The Pyrolysis-Gas Chromatography-Ion Mobility Spectrometer (PY-GC-IMS) combusts, or

pyrolyzes, the biological particles. The biological pyrolysis products are then separated using

gas chromatography. Once separated, the individual pyrolysis products are introduced into an

ion mobility spectrometer for analysis. This technology is still quite new and was developed in a

collaborative effort between Edgewood Chemical Biological Center (ECBC) and the University

of Utah.

The Matrix-Assisted Laser Desorption Ionization-Time of Flight-Mass Spectrometry

(MALDI−TOF−MS) is a variation of mass spectrometry that attempts to use a more gentle

method of ionizing the suspect biological agent than pyrolysis to allow identification of the agent

rather than just broad characterization.

Chemical Biological Mass Spectrometer (CBMS) uses a multistage process to analyze aerosols

for biological content and categorize any biological constituents. The instrument first

concentrates the aerosol, combusts or pyrolyzes it, then introduces the sample into a mass

spectrometer for analysis. An on-board computer is used to analyze the mass spectra for patterns

indicative of biological substances. The instrument is able to categorize biologicals as spores,

cells, or toxins. Figure 5−10 shows an example of the CBMS from Bruker.
29

Figure 5-10. Chemical Biological Mass

Spectrometer (CBMS), Bruker

5.1.4 Identifiers (Specific Identification Technologies)

Identifiers are those components/instruments that are able to identify the suspect biological agent

to the species level (for cellular and viral agents) and toxin type. Specific identification

technologies determine the presence of a specific biological agent by relying on the detection of

a specific biomarker that is unique for that agent. Antibody-based identifiers are used for

systems where speed and automation are required. Where time and manpower are available,

gene-based systems start to take the lead.

The technologies that are used to specifically identify a biological agent are the most critical

components of the detection architecture. These components have the widest variety of

technologies and equipment available. Brief descriptions of several identifiers are included in

sections 5.1.4.1 and 5.1.4.2.

5.1.4.1 Immunoassay Technologies

Immunoassay technologies detect and measure the highly specific binding of antigens

(substances that are foreign to the body) with their corresponding antibodies by forming an

antigen-antibody complex. In an immunoassay-based biological agent identification system, the

presence of an analyte (agent) is detected and identified by relying on the specificity of the

antigen-antibody binding event. The immunoassays are grouped into three categories:

disposable matrix devices (tickets or kits), biosensors that use tag reagents to indirectly measure

binding, and biosensors that do not require a tag (direct affinity assays). Each of these

categories, along with examples of the corresponding technologies, is discussed below.

Disposable matrix devices: Disposable matrix devices are often referred to as tickets or kits.

They usually involve dry reagents, which are reconstituted when a sample is added. There are

one-step assay formats, as well as more complex formats involving multiple steps that are

performed using one or more reagents. Ticket assays can be automated using instrumentation to

perform the manual assay steps and provide a semiquantitative test readout. Rapid handheld
30

assays with greater sensitivity, specificity, and reproducibility are under development for a wide

range of bacterial agents and toxins. These assays have excellent stability characteristics, and

test results are easy to obtain.

Typical ticket-based technologies include the Hand-Held Immunochromatographic Assays

(HHAs), BTATM Test Strips, and the Sensitive Membrane Antigen Rapid Test (SMART) system.

Hand-Held Immunochromatographic Assays (HHAs) are simple, one-time-use devices that are

very similar to the urine test strips used in home pregnancy tests. There are currently 10 live

agent assays in production, four simulants, and five trainers (only saline solution is needed to get

positive results). These tests provide a yes/no response; however, a skilled observer can tell how

much agent is present (semiquantitative measurement) by the degree of color change. HHAs are

currently being used in virtually all fielded military biological detection systems, are in

developmental systems, and are being used by a number of consequence management units.

Their utility is due in large measure to their adaptability to automated readers as well as manual

readers. Power is not required to use HHAs manually.

BTATM Test Strips are detection strips that are manufactured by Tetracore LLC and distributed by

Alexeter Technologies, LLC. The chemistry technique (lateral flow Immunochromatography)

uses monoclonal antibodies that are specifically attracted to the target substance. When the level

of the target substance is present in the sample above a certain concentration, the antibodies and

target substance combine in the BTATM Test Strip to form a reddish band that appears in a

window. The test is positive if two colored lines appear. If only one colored line appears in the

"C" Window, the test is negative. This technique provides fewer false positives in

environmentally collected samples. Anthrax and ricin assays are available, with other assays in

development. Figure 5−11 shows the Tetracore BTATM Test Strip testing procedure.

Figure 5-11. BTATM Test Strip testing procedure, Tetracore, LCC

Sensitive Membrane Antigen Rapid Test (SMART) is a ticket-based system for detecting and

identifying multiple analytes. The core chemistry approach detects antigens in the sample by

immunofocusing colloidal gold-labeled reagents (leveled antibodies) and their corresponding

antigens onto small membranes. Positive results (formation of a red dot) are detected by an

instrument that measures the membrane reflectance. An automated ticket-based system can be

used to perform the SMART immunoassays. Figure 5−12 shows an example of the NDI Smart

Ticket, manufactured by New Horizons Diagnostics Corporation in Columbia, MD.
31

Figure 5-12. NDI Smart Ticket

Reagent Tag Biosensor Approaches: In this approach, biosensors integrate the sensing element

(optical or electronic) with the biological coating to provide for a rapid, simple bio-analysis. In

contrast with tickets, biosensors for biological agent detection consist of a sensing element, often

enclosed in a flow cell, and an associated instrument for quantitative readout. A fluidics system

is required to provide an automated, multi-analyte immunoassay to introduce the sample and one

or more reagents into the sensor/flow cell during a test sequence. Biosensor-based assays are

designed to be automated and often have an inherent capability for multi-analyte detection.

Reagent tag biosensor methods include fluorescent evanescent wave biosensor surface,

electrochemiluminescence, Light Addressable Potentiometric Sensor (LAPS) Immunoassay, and

latex particle agglutination/light scattering.

An example of fluorescent evanescent wave biosensor technology is the Fiber Optic Wave-Guide

(FOWG). The FOWG uses antibody-coated fiber optic probes and a fluorescent “reporter”

antibody to determine the presence of a suspect agent. If an agent is present in the aqueous

solution circulating through the instrument, it will bind to the antibody on the probe. The

instrument then circulates a second solution containing a fluorescent labeled antibody, which

will also bind to the agent. The device then looks for the presence of the fluorescent tag on one

of the probes.

No-Tag Reagent Biosensor Methods: Antigen-antibody binding is detected directly in no-tag

reagent biosensor methods (i.e., direct affinity or homogeneous assays). Advantages to this type

of assay include simplification of the analysis process (fewer steps, fewer components),

minimized disposable fluid use (no need to carry tag reagent solutions), reuse of sensors after a

negative test (minimal disposable use), and a smaller, lighter-weight instrument that consumes

less power.

Examples of no-tag biosensor methods include interferometry, surface plasmon resonance, piezoelectric crystal microbalance, waveguide coupler, and electrical capacitance. The example of

direct affinity no-tag biodetection technology is discussed in the following text. A device that
32

uses no-tag reagent biosensor technology is Bi-Diffractive Grating Coupler (BDG), an optical

transducer that is being developed by Battelle Memorial Institute and Hoffman-LaRoche. This

device takes advantage of a phenomenon linked with one of the two components of a polarized

light wave. Polarized light is divided into a transverse electric (TE) and transverse magnetic

(TM) mode. The TM mode has an evanescent “tail” that moves with the light wave and above

the medium (in this case, a plastic wave-guide that is coated with antibodies specific for a

particular agent). The binding events change the index of refraction of the wave-guide surface

layer, which alters the velocity of light traveling in the wave-guide through its evanescent field

interaction. The optical property measured by this device, using optical interferometry, is the

change in refractive index on the binding of the target molecule with the surface.

5.1.4.2 Nucleic Acid Amplification

Nucleic acid amplification may be used to help detect the presence of DNA or RNA of bacterial

and viral biological agents (nucleic acid amplification cannot directly detect the presence of the

toxins themselves). Samples for nucleic acid analysis can be obtained from field samples, from

laboratory cultures, or from tissues of infected animals or humans. Polymerase chain reaction

(PCR) is the most widely used method to amplify small quantities of DNA for analysis. Two

examples of nucleic acid amplification are included in the following text.

The Mini−PCR (Ten Chamber PCR) is an instrument that has been developed by Lawrence

Livermore National Laboratory (LLNL) and represents one of the first attempts to get genebased identification technologies in a field-useable format. This device relies on a process called

polymerase chain reaction (PCR) and a commercial chemistry called Taq-man®. A suspect

sample is placed into a miniature thermal cycler that heats up and cools off very quickly and has

miniature optics built into it; there are 10 of these mini-thermal cyclers in the 10 chamber device.

In short, the instrument makes many copies of a particular gene segment of the suspect agent (if

the agent is present), and as more copies are made, the more fluorescent light is generated by the

Taq-man® process. The instrument is able to read the increase in light in near real time. This

technology promises to be very sensitive and very specific.

The LightCycler, developed by Idaho Technology, is a thermal cycler that uses a unique builtin fluorimetric detection system with specially developed fluorescent dyes, as well as Taq-man®

technology, for on-line quantitation and amplification products. It is being manufactured under

license by Roche Diagnostics. Figure 5−13 presents a picture of the LightCycler.

The Ruggedized Advanced Pathogen Identification Device (RAPID), from Idaho Technology, is

a rugged, portable field instrument that integrates the LightCycler technology. The RAPID can

run a reaction and automatically analyze the results in less than 30 min. Special software allows

push button use of the RAPID, allowing for quick, safe, and accurate field identification of

possibly dangerous pathogens. It is currently available for military field hospitals and law

enforcement use. See Figure 5−14 for a picture of the RAPID.
33

Figure 5-13. Rapid LightCyclerä,

Idaho Technology

Figure 5-14. RAPID, Idaho Technology

5.2 Standoff Technologies

Standoff systems are designed to detect and identify biological agents at a distance away from

the aerosol/plume or from the detector system, before the agents reach the location of the system.

Standoff systems do not utilize a trigger/cue, collector, or detector but use a bright light source

such as a laser for detection of the biological agents.

Standoff technology uses the concept of detecting and measuring atmospheric properties by laser

remote sensing or LIDAR, an acronym for light detection and ranging. In LIDAR, a short laser

pulse is transmitted through the atmosphere, then a portion of that radiation is reflected back

from a distant target or from atmospheric particles such as molecules, aerosols, clouds, or dust.

All of these systems must be line-of-sight to the suspect biological agent event. Because LIDAR

systems use light, which is composed of short wavelength energy, they are able to “see” the

small aerosol particles characteristic of biological agent attacks (predominantly less than

20 μm in diameter). IR based LIDAR systems are able to see out to ranges of 30 km to 50 km as

the atmosphere is fairly transparent to this wavelength of light. One limiting factor to standoff

systems is the lack of availability of small, inexpensive high-power lasers. Several standoff

instruments are identified below.

IR LIDARs cannot discriminate between biological and nonbiological aerosols; therefore, the

remote detection of biological agents is best accomplished using a UV laser and the laserinduced fluorescence (LIF) technique. This results in an illuminated biological aerosol with a

strong UV laser pulse that causes the biological agent to fluoresce. The fluorescence is redshifted from the UV excitation frequency and detected in a longer wavelength UV band. The

LIF system is more effective during low light or nighttime operations; the range is severely

curtailed by the relative opacity of air to UV light and the high UV background during daylight

hours.
34

Compact LIDAR is a system that has been in development at Soldier Biological and Chemical

Command (SBCCOM) and Edgewood Chemical and Biological Center (ECBC) since 1996. The

goal of the program is to develop a lightweight, ground-based standoff detection system that can

track, calculate relative concentrations, and map potential biological aerosols. The system uses

an IR laser system and cannot discriminate between biological and nonbiological aerosols.

Hybrid LIDAR is a system under development by the Electro Optics Organization Inc. (EOO)

and Stanford Research Institute (SRI), under the sponsorship of the Defense Advanced Research

Projects Agency (DARPA). The goal of this project is to develop a system that can be mounted

on an unmanned aerial vehicle (UAV). The concept is that the UAV will loiter in an area,

scanning for suspicious aerosols with its IR LIDAR component. When a suspect cloud is

spotted, the UAV will move in close and interrogate the cloud for biological content using its

ultraviolet (UV) component.

MIRELA is an IR LIDAR that is being collaboratively developed by SBCCOM and France. The

system was originally developed for standoff detection of chemical clouds but is now being

evaluated for bio-aerosol detection. This system cannot discriminate between biological and

nonbiological aerosols.

MPL 1000 and MPL 2000 are commercially available IR LIDAR systems (manufactured by

Science and Engineering Services, Inc.-SESI) originally developed in collaboration with NASAGoddard Space Flight Center for monitoring atmospheric cloud and aerosol structures. NASA

and DOE now have over a dozen MPL instruments in routine use at research sites. These

instruments cannot discriminate between biological and nonbiological aerosols.

Of the standoff detection systems discussed, the MPL 1000 is the closest to being a fieldable

standoff detection system. The system is already in production and is fairly lightweight and

rugged. This system, as is true with all the systems, requires additional time to develop its

detection algorithm. All of the standoff detection systems described require manual

interpretation of raw data.

The Long-Range Biological Standoff Detection System (LR-BSDS) can detect aerosol clouds up

to 30 km from the detector from an airborne platform, specifically a helicopter. This system uses

pulsed laser beams in the near-IR regime of the optical spectrum (1 μm) to detect these clouds.

However, since only aerosol clouds are detected, there is no biological discrimination to

distinguish these clouds from other clouds, such as dust clouds. See Figure 5−15 for an example

of a long-range detector system.
35

Figure 5-15. Long-Range Biological

Standoff Detection System (LIDARS)

5.3 Passive Standoff Technologies

Passive standoff detection systems rely on the background electromagnetic energy present in the

environment for detection of biological agents. Typically, these systems look at the mid-IR (3 μ

to 5 μ) or far-IR (8 μ to 12 μ) region of the spectrum for agent signatures. Currently researchers

are investigating the utility of IR spectroscopy for detection and identification of biological

agents. While bio-aerosols have been visualized by IR systems immediately after dissemination,

they quickly loose that signature and become invisible to current passive systems. Systems such

as the M21 Remote Sensing Chemical Agent Alarm (RSCAAL) and Joint Service Lightweight

Standoff Chemical Agent Detector (JSLSCAD) have been used in attempts to detect biological

agents with little success.
37

6. HOW TO PREPARE FOR A BIOLOGICAL INCIDENT

This section provides emergency first responders and other interested organizations with

information on what actions an emergency first responder should take in the case of a biological

incident. It has information on Federal and State programs for support, crisis management, and

functional tasks during a terrorist attack.

6.1 Federal and State Programs for Support

As outlined in previous sections of this guide, biological detection equipment is mainly in the

developmental phase. Because of this, there is a limited number of commercially available

instruments; what is available is costly and has limited utility. Without equipment to detect and

identify a biological agent, emergency first responders must turn to existing State and Federal

organizations for support.

A number of State and Federal agencies are working throughout the country to set up standards

for operations during a terrorist attack involving biological, chemical, or nuclear weapons of

mass destruction. The Centers for Disease Control and Prevention (CDC) is coordinating a

nationwide program called the National Laboratory System (NLS) to provide communication,

coordination, and testing capacity required to effectively detect and report disease outbreaks and

exposures (see app. B, ref. 1). The goal of the NLS is to integrate the reporting and response of

disease outbreaks and/or terrorist bioweapon attacks. The system will integrate Federal, State,

and local public health laboratories, as well as hospital, independent, and physicians’

laboratories, for monitoring the population for an outbreak of disease. Through the NLS, the

CDC provides private and State public health laboratories with information, analytical methods,

and analytical reagents for analysis of biological agents. The CDC also sponsors the Laboratory

Response Network (LRN) through the Association of Public Health Laboratories (see app. B, ref.

2). The LRN is focused on educating laboratories on the methods needed to test for biological

agents.

The emergency first responder must recognize that while public health laboratories and

supporting clinical laboratories have the capability to detect and identify possible biological

agents, these tests are not field deployable. The detection methods used are laboratory-based

systems and should not be confused with field-based systems described in earlier portions of this

guide. Generally, the laboratory-based systems are slower than field systems, but the laboratorybased systems exhibit greater selectivity and versatility than field-based systems. It should also

be recognized that different laboratories have different capabilities.

The CDC uses a four-level categorization of laboratory responsibilities for detection and

identification of a biological agent. The laboratories are categorized as Level A, Level B, Level

C, and Level D laboratories.

• Level A laboratories focus on early detection of intentional dissemination of biological

agents. They are mostly composed of microbiology laboratories that conduct primary

clinical testing, such as hospital and independent laboratories. Level A laboratories are
38

responsible for ruling out the presence of pathogenic organisms and forwarding suspicious

and potentially dangerous organisms to laboratories capable of identifying the organisms.

• Level B laboratories focus on testing for specific agents and forwarding organisms or

specimens to higher level biocontaminant laboratories.

• Level C laboratories focus on advanced and specialized testing for rapid identification of

biological agents.

• Level D laboratories focus on diagnosis of rare and dangerous biological agents.

First responders will generally only have to deal with Level A laboratories.

6.2 Crisis Management in a Terrorist Attack

Crisis management must be integrated and managed under an overall unified command structure

during a terrorist attack (see app. B, ref. 3). Crisis management for a terrorist attack using

biological agents consists of public health monitoring, surveillance, detection, and reporting the

use of a biological weapon of mass destruction (WMD).

Emergency first responders (fire and rescue) will be involved in the early stages of crisis

management, primarily the reporting of the possible use of a biological weapon. For this reason,

emergency first responders need to have an emergency response plan in place for any possible

biological (as well as chemical and radiological) incident. Therefore, it is strongly recommended

that emergency first responders plan their response to a biological (as well as chemical and

radiological) incident well in advance.

A recent report in the State of Maryland entitled “Maryland Health and Medical System

Preparedness and Response Plan––Weapons of Mass Destruction, Work Plan,” suggests that the

response to an incident be coordinated through local, State, and Federal channels to ensure

complete integration of the local response to any such incident (see app. B, ref. 3). The State of

Maryland recommends coordination with the State police, the State public health

department/laboratories, and the Federal Bureau of Investigation (FBI). It is stressed that these

are the recommendations of the State of Maryland; recommendations may be different for each

State. Therefore, it is essential that the first responder contact local and State officials in order to

coordinate a response to a biological agent incident.

6.3 Functional Tasks During a Terrorist Attack

In the event of a terrorist attack using biological agents, each supporting agency has different

functional tasks that must be carried out. Local fire and rescue service’s functional tasks state

that, “The Fire Chief, or first ranking officer on the scene, will be the initial incident commander

for single point source incidents and must make initial determinations on tactical responses and

additional support ....” (see app. B, ref. 3). Local officials must plan ahead for this contingency

by providing senior officers of the fire and police departments with education and training on the

identification of biological (and chemical or nuclear) incident.

Once it is determined that the event is a result of a release of a biological agent (either by a

terrorist or accidental), the appropriate authorities must be contacted. In the State of Maryland,

first responders should contact the Maryland State Police who are to “assist with early detection
39

and monitoring activities by notifying the Department of Health and Mental Hygiene and the

Local Health Officer of threats, credible threats, impending events, or actual terrorist acts that

may produce casualties” (see app. B, ref. 3). Each first responder unit must first determine the

response chain for their particular State. In this way, the first responder is integrated into the

overall response to a biological (and chemical or nuclear) incident.
41

7. SUMMARY

An Introduction to Biological Agent Detection Equipment for Emergency First Responders was

developed to provide information to the emergency first responder community and aid their

understanding of biological agent detection equipment. Information included in the guide

focuses on biological agents, challenges of detection, components of detection, and the basic

technologies that have been or are being considered in the research and development (R&D) of

biological agent detection equipment.

The guide identifies a number of biological agent detection technologies and some equipment

associated with the technologies.

5

It is important to note that the equipment referenced is not all

inclusive with what is currently available or currently being tested. While some equipment is

commercially available, most is not (a notable exception is Tetracore test strips for biological

agents).

6

It is also important to realize that biological detection equipment is limited with

respect to biological agents detected as well as operational conditions. Because of this, An

Introduction to Biological Agent Detection Equipment for Emergency First Responders was

written to serve the first responder community as a guide to the status of biological agent

detection.

Because commercially available biological agent detection equipment prices range from tens to

hundreds of thousands of dollars, it is obvious that R&D efforts will have to continue.7 These

efforts will focus on lowering equipment costs while improving equipment sensitivity and

selectivity. As new equipment and technologies emerge, and more importantly for the first

responders, as equipment becomes commercially available, this guide will be updated.

Because of the lack of affordable detection equipment for biological agents, first responders must

integrate their response into the overall national effort. This national effort is being developed

by the CDC as well as the FBI and includes the development of analytical assets at State health

laboratories for detecting biological agents. The link from the first responders to the national

response effort is most likely the State police and the State public health laboratories. However,

this plan is based on the State of Maryland plan and may be different for each State. Therefore,

in developing a response plan for biological weapons, it is recommended that first responders

contact their State police to determine if a standard operating procedure (SOP) for a terrorist

attack using biological, chemical, or nuclear WMD exists. It is also suggested that prior to an

event involving a biological WMD, first responders contact the nearest public health laboratory

to determine points of contact. Appendix B lists the phone numbers for public health

laboratories in most States, as well as the Association of Public Health Laboratories (a nonprofit

association working to actively promote the interest of public health laboratories), and internet

addresses for the Association of Public Health Laboratories, CDC, and State Public Health

Laboratory home pages (see app. B, ref. 2, 4, and 5).

 

5

It is critical to understand that reference to these technologies and equipment by trade name, trademark, manufacturer, or otherwise does not

necessarily constitute or imply its endorsement, recommendations, or favoring by the United States Government.

6

For example, immunoassay tickets are relatively inexpensive; however, the antibodies that are required for identification of the biological agents

are not commercially available.
APPENDIX A––REFERENCES
A−1

APPENDIX A––REFERENCES

1. John A. Barrett, Gregory W. Bowen, Scott M. Golly, Christopher Hawley,

William M. Jackson, Leo Laughlin, Megan E. Lynch, Assessment of Biological Agent

Detection Equipment for Emergency Responders, June 1, 1998. Chemical Biological

Information Analysis Center (CBIAC), P.O. Box 196, Gunpowder, MD 21010-0196.

2. Chemical and Biological Terrorism: Research and Development to Improve Civilian

Medical Response to Chemical and Biological Terrorism Incidents, National

Academy of Sciences, 1999. National Academy Press, 2101 Constitution Avenue,

N.W., Box 285, Washington, DC 20055.

3. John A. Barrett, Gregory W. Bowen, Scott M. Golly, Christopher Hawley, William

M. Jackson, Leo Laughlin, Megan E. Lynch, Final Report on the Assessment of

Biological Agent Detection Equipment for Emergency Responders, U.S. Army

Chemical and Biological Defense Command (CBDCOM), June 1, 1998. CBIAC,

P.O. Box 196, Gunpowder, MD 21010-0196.

4. Assessment of Biological Warfare Detection (CD) Joint Program Office for BioDefense, Skyline #2, 5203 Leesburg Pike, Suite 1609, Falls Church, VA 22041-3203,

September 13, 1999.

5. State of the Art Report on Biodetection Technologies, July 1995. CBIAC, P.O. Box

196, Gunpowder, MD 21010-0196.

6. B. Newman, “Opening the Case of the Poison Umbrella,” The Wall Street Journal,

May 24, 1991. E-mail address:

http://chiron.valdosta.edu/rgoddard/biol4900/corbett/corbett.htm .
APPENDIX B—CONTACT INFORMATION

FOR FIRST RESPONDERS
B−1

Telephone Numbers for State Public Health Laboratories

Association of Public Health Labs 202−822−5227

Alaska 907−269−7942

Arizona 602−542−1194

California 510−540−2408

Colorado 303−692−3289

Connecticut 860−509−8540

Florida 850−245−4401

e-mail Bill Dart (Bill_Dart@doh.state.fl.us)

Georgia 404−327−7900

Idaho 208-334−5939

Illinois 217−782−4977

Indiana 317−233−8006

Kansas 785−296−1620

Louisiana 504−568−5375

Maine 207−287−2727

Massachusetts 617−983−6200

Michigan 517−335−8063

Minnesota 651−215−5800

Missouri 573−751−0633

Nebraska 402−552−3350

New Jersey 609−292−0430

New Mexico 505−841−2500

New York 716−898−6100

North Carolina 919−733−7834

Ohio 888−634−5227

Oklahoma 405−271−5070

Oregon 503−229−5882

South Dakota 800−738−2301

Tennessee 615−262−6300

Texas 512−458−7228

512−458−7676

Utah 801−538−6128

Vermont 802−863−7240

800−640−4374

Virginia 804−786−7905

Washington 206−361−2800

West Virginia 304−558−3530

Wisconsin 888−494−4324

Wyoming 307−777−7431
B−2

Suggested Websites and Addresses for More Complete Information

1. National Laboratory System (NLS) Division of Laboratory Systems (DLS):

http://www.phppo.cdc.gov/mlp/nls.asp .

2. Association of Public Health Laboratories: http://www.aphl.org/ .

3. Maryland Health and Medical System Preparedness and Response Plan––

Weapons of Mass Destruction, Work Plan, James R. Stanton, Maryland Institute

for Emergency Medical Services Systems (410-706-0415), May 2000.

4. Center for Disease Control: http://www.cdc.gov/ .

5. Public Health Laboratory listings:

http://www.phppo.cdc.gov/DLS/links/links_phl.asp .
ABOUT THE LAW ENFORCEMENT AND CORRECTIONS

STANDARDS AND TESTING PROGRAM

The Law Enforcement and Corrections Standards and Testing Program is sponsored by the Office of

Science and Technology of the National Institute of Justice (NIJ), U.S. Department of Justice. The program

responds to the mandate of the Justice System Improvement Act of 1979, directed NIJ to encourage

research and development to improve the criminal justice system and to disseminate the results to Federal,

State, and local agencies.

The Law Enforcement and Corrections Standards and Testing Program is an applied research effort that

determines the technological needs of justice system agencies, sets minimum performance standards for

specific devices, tests commercially available equipment against those standards, and disseminates the

standards and the test results to criminal justice agencies nationally and internationally.

The program operates through:

The Law Enforcement and Corrections Technology Advisory Council (LECTAC), consisting of

nationally recognized criminal justice practitioners from Federal, State, and local agencies, which assesses

technological needs and sets priorities for research programs and items to be evaluated and tested.

The Office of Law Enforcement Standards (OLES) at the National Institute of Standards and Technology,

which develops voluntary national performance standards for compliance testing to ensure that individual

items of equipment are suitable for use by criminal justice agencies. The standards are based upon laboratory

testing and evaluation of representative samples of each item of equipment to determine the key attributes,

develop test methods, and establish minimum performance requirements for each essential attribute. In

addition to the highly technical standards, OLES also produces technical reports and user guidelines that

explain in nontechnical terms the capabilities of available equipment.

The National Law Enforcement and Corrections Technology Center (NLECTC), operated by a grantee,

which supervises a national compliance testing program conducted by independent laboratories. The

standards developed by OLES serve as performance benchmarks against which commercial equipment is

measured. The facilities, personnel, and testing capabilities of the independent laboratories are evaluated by

OLES prior to testing each item of equipment, and OLES helps the NLECTC staff review and analyze data.

Test results are published in Equipment Performance Reports designed to help justice system procurement

officials make informed purchasing decisions.

Publications are available at no charge through the National Law Enforcement and Corrections

Technology Center. Some documents are also available online through the Internet/World Wide Web. To

request a document or additional information, call 800–248–2742 or 301–519–5060, or write:

National Law Enforcement and Corrections Technology Center

P.O. Box 1160

Rockville, MD 20849–1160

E-Mail: asknlectc@nlectc.org

World Wide Web address: http://www.nlectc.org

This document is not intended to create, does not create, and may not be relied upon to create any rights,

substantive or procedural, enforceable at law by any party in any matter civil or criminal.

Opinions or points of view expressed in this document represent a consensus of the authors and do not

represent the official position or policies of the U.S. Department of Justice. The products and manufacturers

discussed in this document are presented for informational purposes only and do not constitute product

approval or endorsement by the U.S. Department of Justice.

The National Institute of Justice is a component of the Office of Justice Programs, which also includes the Bureau of Justice

Assistance, the Bureau of Justice Statistics, the Office of Juvenile Justice and Delinquency Prevention, and the Office for Victims

of Crime.