Detection Methods in Rapidly Deployable Laboratories for CBRN Emergencies

Royal Higher Institute for Defense – 12th March 14

Pr. Dr. JL GalaMed Colonel,

Director BE Mil-AcadBiotech Platform CTMA/DLD-Bio

Center for Applied Molecular Technologies /

Defence Laboratories Department-Biothreats

UCL-Brussels, Belgium

OBJECTIVE

� Genetic diseases

� Biomarkers / Biosensors

� Infectious diseases :

identification of

pathogens, virulence &

antibiotic resistance…

Academic (UCL) & Clinical pole

(St Luc Univ Hospital)

BE-Defence(DLD)

RESOURCES

Applied sciences - Technological transfer unit

ClinicalGenetic Testing

DLD-Bio

Genetic Research

Spin-off

Identification

Diagnosis

1

Engineering

Biostatistics

Bioinformatics

Biological –Medical genetics

CTMA / DLD-Bio

R&D Clinical / Ops applicationsTransfert

Also in the field !

AIM: To connect EU projects dealing with deployable capacities !

1. FP7- SEC-MIRACLE (2013): MobIle Laboratory for the Rapid Assessment of CBRN Threats Located within and outside the EU (CTMA coordination)

2. FP7-SEC-PRACTICE (2011): Preparedness and Resilience against CBRN Terrorism using Integrated Concepts and Equipment

3. FP7-SEC-EDEN (2013): End-user driven DEmo for CBRNE

4. FP7-SEC-CAERUS (2014): Evidence-based policy for post-crisis stabilisation:bridging the gap

1. EBLN-EMS4 - Database of B-agents (Genetic profiling of deadly B-agents)2. JIP-CBRN call-1 & -2: - BFREE (CBRN mixed samples

handling and analysis) - RACED (monitoring of

decontamination

EFC

B-LiFE - IAP/ARTES20: Biological Light Fieldablelaboratory for Emergencies(CTMA coordination)

ESA

EDA

EC-FP7-SEC (CBRN)

CBRN matrix

(a) « CBRN emergencies » in our daily life ?

(b) Most worrying threats ?

(c) Rationale for rapidly deployable capacities and

related issues ?

Detection Methods in Rapidly Deployable Laboratories for CBRN Emergencies

(a) What do CBRN(E) threats mean in our daily life ?

6

March 1995

Augustus 2013

Lethal coumpoundsNeed for rapid local identification – Forensic aspect !

March 1988

7

Nov 2006

Nov 2004

?

Copyright 2012 - CTMA/DLD-Bio

Death rate : 50- 70%

Contagious and lethal diseasesNeed for rapid local management !

Viral Hemorraghic Fevers in Africa :

3 different outbreaks in 2012 !

Anthrax outside «military field» !

CTMA/ DLDL-Bio

? ?

Deadly anthrax cases (USA Sept 2001)

versus

Countless anthrax copycats

White powder – Anthrax

BELGACOM towers, Belgium, 11 Jan 2013

Deliberate release

human infection ?

?

(a) What do CBRN threats mean in our daily life ?

(b) What are the most worrying CBRN threats ?

(c) Rationale for rapidly deployable capacities and

related issues ?

Detection Methods in Rapidly Deployable Laboratories for CBRN Emergencies

annéesheuresHours Days Years

De

ath

s

C / E / R

BC / R / E

B

Biological agents :

Mostly covert incidents with delayed lethality

Impact = (% affected) X (speed of spread)

Risk = Likelihood X Impact

Common Mil-Civ Needs:Joint efforts and cooperation

VIRUSES FUNGI PROTOZOAEBACTERIA PRIONS (?)

Challenges: Rapid, specific and sensitive DIM

- Deadly for human beings, animals and crops- Contaminating the environment

TOXINS

Mitigate CBRN risks : same identification tools and technologies : Dual !

Intentional release onlyCBRN cases

Natural outbreak –Endemic zone

Daily infectionsHospital – Ops «in-field»

POTENTIAL THREATS OCCASIONAL THREATSCOMMON THREATS

Best validation & trg model

NOT EXPECTED in environment

Civ-Mil Concern

Terrorism - Risk in war zone (Ops)

PRESENT in natural environment

Civ-Mil Concern

Risk in Epid & endemic area in Ops

COMMON in daily life

Civ-Mil Risk anytime, anywhere

Detection of Biological Warfare Agents (BWA): The "Dirty Dozen" (WHO)

Initial Non SpecificSymptoms

Pathogen

Bacteria

FLU SYNDROME

NONSPECIFIC SYMPTOMS

[fatigue, myalgia, fever, headache, nonproductive

cough, chest pain, respiratory distress, high

fever, abdominal pain vomiting, diarrhea]

Inhalation AnthraxBacillus anthracis

Pneumonic PlagueYersinia pestis

TularemiaFrancisella tularensis

(Rabbit Fever)

BrucellosisBrucella suis

Q-FeverCoxiella burnetii

Glanders:Burkholderia mallei

Burkholderia pseudomallei

Viruses SmallpoxVariolavirus

Venet. Equine EnzephalitisVEE-Virus

Marburg-Fever(Viral Hemorrh. Fever)

Marburg-Virus

Toxins BotulismClostridium botulinum

Ricin-IntoxicationRicin

SEB-IntoxicationStaphylococcal-Toxin

STANAG 4632

(Edition 1)

Study Draft 3

NAVY/ARMY/AIR

NATO STANDARDIZATION AGREEMENT

(STANAG)

DEPLOYABLE CBRN Analytical laboratory

STANAG 4632

CDC Cat. Agent Name

A Bacillus anthracis (Anthrax)

Yersinia pestis (Plague)

Francisella tularensis

Smallpox

Viral hemorrhagic fevers : filoviruses and arenaviruses

Botulinum Toxin

B Brucella melitensis

Burkholderia mallei

Burkholderia pseudomallei

Coxiella burnettii

encephalitis viruses (e.g. VEE, EEE-virus)

Chlamydia psittaci

Rickettsia prowazekii

Ricin toxin

Staphylococcal enterotoxin B

Toxins of Clostridium perfringens (alpha, beta,

epsilon and iota)Food and water safety

threats

(a) What do CBRN threats mean in our daily life ?

(b) What are the most worrying CBRN threats ?

(c) Rationale for rapidly deployable capacities and

related issues ?

(1.Lab architecture, 2.Sample collection, 3.Type of sample, 4.DIM,

5.results interpretation, 6.logistical constraints)

Detection Methods in Rapidly Deployable Laboratories for CBRN Emergencies

Rationale: Some agents are very contagiousrapidly lethal, no reliable clinical identification

Monkeypox, Chickenpox… or Smallpox ?Need for rapid differential diagnostic tests !

Protect RecoverRespond

Prevent

Deter

Act

ions

to

unde

rtak

eag

ains

t

caus

es

Act

ions

to u

nder

take

agai

nst

effe

cts

Before incident During After incident

PREPAREDNESS / PLANNING

RESPONSE / RECOVERY

DETECTION - MONITORING - IDENTIFICATION

SITUATIONAL AWARENESS & RESILIENCE

Rationale for radidly deployable capacity ?Role in the whole CBRN Disaster Life Cycle

CB

RN

Inci

dent

-C

onse

quen

ces

-C

risis

Man

agem

ent

Dec

isio

nS

uppo

rt -

Com

mun

icat

ion

To reinforce EU preparedness to crises affecting internal-and external-borders

1. What is the best architecture for a CBRN health crisis

Mobile capacityfor CRBN or for Health crisis ?

CBRN crisis

MILITARY specificationsCIV-MIL specifications

Health crisis

Scarce « Ops-certified » structure – funding?Several « Ops-certified » structure – joint capacity is possible – funding possible !!!

Mobile capacityfor CRBN or for Health crisis ?

Common Civ-Mil specifications– HARDWARE: Analytical equipement– SOFTWARE: Communication, site selection, georeferencing– HUMAN RESOURCE: Expertise

– Full CBRN shelter protection – Heavy shelter, slowly deployable– Sampling team– Ops military C2

– B-specific– Light Fieldable– First responders (nurses, MD)– GOARN / WHO / EC DEVCO

CBRN crisis

MILITARY specificationsCIV-MIL specifications

Common HEART

Health crisis

From a simple inflatable tent … To a wooden hut…

Extremely rapidly deployable and Ops !

Gary Kobinger (PHA CA,

Personal Communication)

Kuya Kumpala Camp Kampungu Lab Layout

CBRNE CoE, Square Meeting, Brussels

2. On site sample collectionDuring / After a CBRN health

crisis ?

Triage and

decontamination

Casualty clearing

station

Ambulance

loading point

Outer cordon

Out

In

Incident control

point

Silver

command

Inner cordonIncident

Hot zone Warm zone Cold zone

Clean/dirty line

PPE

CBRN Incident

For CBRN crisis due to lethal B-agents:

Aerosol spread – critical infrastructure

Deadly outbreak - Health crisis: Limits ?

Military Sampling team : EU-Security and Research

Conference (SRC'10), Oostende, Sept 2010

Belgian presidency of the EU–council

14 Cie – 4 BnGn, Jambes, BE-Defence: Processing of a suspected sample(manual or automated collection, decontamination, transport to the field lab)

Civilian sampling team for major

natural outbreaks

Viral Hemorraghic Fever

3. What type of samples ? … Any type !

Sampling for a quick analysis of risks: defined by the type of CBRN

crisis

Environmentalsamples

Various Matrixes: Air / Soil / ground

Water / Crops

National team of experts

NATO/SIBCRA

Biologicalsamples

(Human / animal)

Various Matrixes: Blood, aspirate(CSF, lung),feces, urine, tissue biopsy…

Med / Vet team

?

Point of care testing

4. DIM Methods ( Detection, Identification, M onitoring) ?

DIM or the quest for the « Holy Grail »

The « Too Many » issue !

Many…… • Actors and Disciplines• Objectives • Pathways: wind, climate, matrixes…• Targets (strains,…)

– Background little understood – Pathogenic strains difficult to distinguish from near

neighbours

• But also many methods !(qualitative or quantitative…)

In-field Genetic-based identification: Rapid, specific, reproducible, safe !

Lung infection

Daily hospital challenge : Detect to treat !

…..Need for rapid identification tools

Various causes (terrorism – environnement – hospital…..) – common symptoms

Bone infectionSkin infection

In-field monitoring of health-threateningagents nearby a Military settlement

• Monkeypox outbreak

• Ebola outbreak

Real-time PCR

• Resistant Tuberculosis

Endpoint-PCR

Response time

Selectivity Sensitivity Warning Detection Identifi-cation

Monito-ring

Survey / recce

MolecularGenetics ☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺ --- + +++ + +

Immuno ☺☺☺☺ ���� ���� - ++ - + -

Opticaldetection ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺ ? ☺☺☺☺☺☺☺☺ ? +++ ☺☺☺☺☺☺☺☺ ? ☺☺☺☺☺☺☺☺ ? + +

Physicaldetection ☺☺☺☺☺☺☺☺ ���� ���� ++ + - ++ +

HybridTechnologies ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺ ☺☺☺☺☺☺☺☺☺☺☺☺

Holy Grail: hybrid identification methods !

Holy Grail

5. Results Interpretation and reporting

Standardisation of Methods –Harmonization of procedures

• Specificity/selectivity• Sensitivity / Limit of detection (LOD)• Precision (within the laboratory repeatability and/or

within the laboratory reproducibility conditions)• Uncertainty of measurement (e.g., 95% CI)• Linearity and working range• Accuracy (bias) (under within laboratory

repeatability and/or within laboratory reproducibility conditions) at the threshold concentration

• Precision (under within laboratory repeatability and/or within laboratory reproducibility conditions) at the threshold concentration

• Stability, Ruggedness and Robustness• False-alarm rate• …….

- Internal Validation

- Proficiency tests

(round robin )

Internat. Org. for

Standard.

(ISO 17025, 15189…)

EC-JRC policy

In Bio Weaknesses !!!

6. Logistical constraints

Deployment constraints

• Balance the volume/weight of the equipments with the technical /logistic needs and constraints

• Robust /portable “cutting edge” technologies: avoid the loss or damage of often sensitive equipments during the journey and deployment.

• Selective, sensitive, reliable and validated identification of health-threatening B-agents in a large range of samples (complex matrixes)

• Compliant with Biosecurity and Biosafety standards

Workers health protection: Biosafety level: BSL4 - BSL3- BSL2 ?

… Anyhow always « inactivate » the sample!

Gary Kobinger (PHA CA, Personal Communication)

- Preliminary inactivation of biological agents

- Nucleic acid based-identification

Miniaturisation – Ruggedization

Do-it-yourself MOTS

Prevention

B-agent inactivation on the

field

Miniaturisation

Rhodotron Accelerators

INDUSTRIAL ELECTRON-BEAMPROCESSING

Main issue: Unbroken cold chain

Lyophilization of the reagents !

6. Way ahead

44

B-Life Projects: Integration of Space-based technologies Sat Communications, Sat Navigation (GPS/GNSS), Earth observation (site selection)

GPS – EGNOS – GALILEO

Georeferencing Localization of samples

& Sampling team Localization

Copyright 2012 - CTMA/DLD-Bio

Earth Observation: (a) Site selection

Candidate deployment

sites 50-500m

(b) Sampling team guidance

Sat- and phone-guidance

(c) Crisis Evaluation

Population estimations, trends

and monitoring over time

Al Mafraq – Jordan

Refugee camp

authorized area

Integrate Multidisciplinary

CBRN expertise

Avoid CBRN fragmentation

Identification (assays)Provisional, unambiguous, Forensics

CBRN (clinical/environmental)

samples in the field

Biosafety issues(inactivation , and decontamination)

Crisis intensitySingle >< Mixed CBRN threats

Pre-analytical

Analytical

Standardisation(Interoperability, Procedures, Equipments)

Inside >< outside EUCross-border crisis

� EU Integration (existing EU

initiatives and capacities :

SANCO, ECDC, ECHO,…)

� Compatibility with existing

international expertise

� Sustainability (business case)

� Evolution (Cutting edge

technologies, training, skills,

knowledge)

Overhead structure or organisation

Biosecurity - Logistics - Secure Communication / Interoperability - Crisis management

EC/SEC/MIRACLE PROJECT

CBRN mobile lab : Definition? Need?

…… Global Architecture ?

Scalability

END