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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Weiter- und Fortbildungs-Symposium
Molekulare Diagnostik 5. März 2015
Dr. Marc Strasser, Head Virology, SPIEZ LABORATORY
Ebola outbreak
and the SPIEZ
LABORATORY
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
• The Ebola Virus
• Transmission
• Ebola outbreak - current
situation in westafrica
• Control strategies
• Tasks of SPIEZ LABORATORY
• SPIEZ LAB field mission (EmLab) in Westafrica
Content
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The Ebola Virus
• Filovirus (Filoviridae)
• Variable relevant spezies (Ebola, Marburg, Reston)
• Incubation time: 2 - 21 days
• Letality: 30 - 90%
• Transmission: through body fluids
• Symptoms: diarrhea, vomiting, head- and bodypains, fever,
rash, inner and external bleedings
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The Ebola Virus
Properties:
• Filoviruses are filamentous
Viruses (latin: filum)
• Envelopped,
single stranded-RNA-Viruses.
• Length between 1‘000 and 14‘000 nm, dimeter 60 to 80 nm
• 1967: First cases of Filovirus (Marburg) in Europe.
• 1976: First Ebola outbreak (Ebola River in what is now the
Democratic Republic of the Congo)
Befällt die meisten Zellen eines Organismus
100 nm
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The Ebola Virus
Replikation cycle:
May infect and propagate in most of the human tissue
100 nm
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The Ebola Virus
• Filovirus (Filoviridae)
• Variable „human relevant“ spezies (Ebola, Marburg)
• Incubation time: 2 - 21 days
• Fatally rate (lethality): 30 - 90%
• Transmission: through body fluids
• Symptoms: diarrhea, vomiting, head- and bodypains, fever,
rash, inner and external bleedings
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The Ebola Virus
• 2007
Uganda
• 37 deaths
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The Ebola Virus
• Filovirus (Filoviridae)
• Variable relevant spezies (Ebola, Marburg, Reston)
• Incubation time: 2 - 21 days
• Letality: 30 - 90%
• Transmission: through body fluids (actually !!)
• Symptoms: diarrhea, vomiting, head- and body pains, fever,
rash, inner- and external bleedings
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Transmission (Reservoir)
• Fruit bats (Rousettus aegyptiacus) ,
living in the tropical rain forest are in
fact the reservoir for the Ebola Virus
(published in 2007).
• They carry the pathogen without
getting ill.
• Fruit bats segregate the pathogen
through urine and saliva.
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Transmission (other Spezies)
• Fruit bats are licking and chewing
fruits and remains of them will be
splitted out.
• Such scraps of fruit are eaten by eg.
primates, antilopes and boars.
• Transmission and infection occurs
through little lesions and wounds in
mouth or skin of these animals.
• Mostly of such infected animals get
seriousely ill and subsequently die.
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Transmission (by Bushmeet)
• Il or dead animals (bushmeet)
are collected by the poor
population, cutted in multiple
bloody pieces and shared with
family or clan members.
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Transmission (rituals)
• Rituals are very widespread
in the westafrican region.
• Ill persons or dead bodies
are touched, washed, kissed
and embrassed without any
protection.
• This mode of behaviour
promotes the transmission of
the Ebola Virus through
body- and fluid contact and
represent therefore a
huge risk of infection.
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Transmission (Cycle)
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Ebola outbreak in Africa
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Ebola outbreak in Africa
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Distribution in 2014
June July August
Ebola outbreak in Africa
The indicator case was already registered in September
2013 but not recognized as an Ebola infection.
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Situation as of February 2015
Source: CDC, Atlanta, USA
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Situation ausserhalb Westafrikas
6.Oktober 2014
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Total cases February 2015
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Guinea (66.7% )
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Sierra Leone (29%)
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Liberia (42%)
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
First isolates from Ebolastrain Zaire (Guinea)
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Marburg outbreak
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Control strategies
National (in west africa):
• Build up of medical hospital wards.
• General education concerning the disease
• Build up medical care
• Military operation
International missions:
• Medical care people (MSF)
• Build up Laboratory capacity (eg. EmLab)
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Current Countermeasure strategies
Vaccines:
Multiple variants are succsessfully tested on NHP ‘s.
Two of them are actually tested on volunters in Switzerland (HUG,
CHUV).
• In future vaccines can be applied prophylactic.
• The application must be comprehensiv (a whole population
should be vaccinated)
Antibodies:
Artificially produced antibodies (eg. ZMapp) or reconvalescent
serum (from survivors) are promising if applied in an early stage of
infection.
Antiviral Substances:
Existing effective chemical substances against viral infections (eg.
Interferon, Tamiflu, Ribavirin).
• Antibodies and antivirals may be used in an early stage of
infection (as soon as possible).
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Vaccine trials 1
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Vaccine Trials 2
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Tasks of SPIEZ LABORATORY
Biocontainment Laboratory
BSL-4
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
SPIEZ LABORATORY
Swiss Institute for NBC Protection and NBC Therats.
Mission of the biology branche (1995 – to date):
Detection of special highly pathogenic human microorganisms
and toxins as well as detection of potentially warefare Agens
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Birds view
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Organigram
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Biology branch: Main Functions and Tasks
Diagnostics
- Primary diagnostics for RG3, RG4
- Differential diagnostics
- Analysis of unknown samples → CBRN
Research
- Basic and applied science
Training
- Provider for Biosafety / Biosecurity
Reference Function
- WHO Collaborating Center (Biosafety)
- National Reference Center VHF
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Biology
Bakteriology
Virology
Toxinology
- Watercontrol for the
Swiss Army
- Scientific Projects
with external Partners
- Detection and
diagnostics of RG 3
and RG 4 viruses
- Detection and
diagnostics of RG 3
bacteria
- Detection and
diagnostics of
biological toxins
- Diagnostics in
molecular biology
- Development and
validation of methods
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Project Biocontainment Laboratory
1996 Initiation of the Project
2001 Site Selection
2003 Political Commitment (DDPS)
2005 Enviromental Impact Study > Building permit
2006 Submit Project to Parliament > Financial Support
2005 (28.5 Mio CHF)
2007 Start Construction
2012 Initial Operation
(BSL-2, BSL-3 Labs)
2012 – 2013 Integral testing and
validation
2014 Operation of the BSL-4 Lab
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Risk groups and Biosafety levels
RG 1: no risk (Bakers yeast) → BSL 1
RG 2: slight risk (Salmonella) → BSL 2
RG 3: moderate risk (Anthrax) → BSL 3
RG 4: high risk (Ebola, Smallpox) → BSL 4
Risk groups RG Biosafety Levels BSL for microorganisms for Laboratories
Biocontainment laboratory Spiez
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The biocontainment laboratory allows the safe handling of pathogenic micro-
organisms in the highest risk groups (RG 3 and 4).
Safety is the number one priority in the biocontainment laboratory to the aim
that the public and the environment are protected from release of infectious
pathogens and that the staff is protected from lab associated infection.
Biosafety related tasks include:
• Guaranteeing biological safety
• Guaranteeing technical safety
• Training
Biocontainment Laboratory
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Containment Concept
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Level 4 Laboratory
Primary Barriers Secundary Barriers
All works in MSC III
or in MSC I + II benches in
Combination with protective
Suits under overperssure
> Protection of Staff
- tight Construction
- Locker Systems
- Double door access
- Underpressure
- Air filtration (HEPA)
> Protection of the Enviroment
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Biocontainment Laboratory: Design
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
The lab staff is protected from the pathogens
Biocontainment Laboratory
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Society / environment is protected from pathogens
Filtration of incomig and outgoing air
using HEPA filters (H14)
Waste water treatment (sterilisation)
Biocontainment Laboratory
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Inactivation of patient samples
in BSL3+ (Glovebox) or BSL4 lab
• Inactivation of whole blood & serum in
extraction reagent (AVL-buffer)
• Sluice out inactivated material
• time required: 2-3 h
RNA extraction: time required: 1 h
Mastermix and amplification RT-PCR:
• time required: 2 h
Time for total primary diagnostic approx 6 h
Confirmatory tests: ->
Ebola Virus Diagnostic: Laboratory Requirements
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
End-point RT-PCR with sequence analysis
• time required 2 days
Virus isolation in cell cultures
• from whole blood, serum, urine,
clinical and environmental swabs
• time required 7-9 days
Further characterization:
• Whole genome sequencing
->Methods are being established
• Electron microscopy
Ebola Virus Diagnostic: Confirmation Tests &
further analysis
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Objective:
Dose finding, safety and
immunogenicity phase I/II vaccine
study
• Recombinant Vesicular Stomatitis
Virus in which envelope
glycoprotein (GP) is replaced by
Zaire Ebola virus GP.
• In vivo replication is limited in
NHP, but release of large amounts
of ZEBOV GP.
• No serious adverse effects
observed with VSV-ZEBOV
Ebola vaccine phase l/II trial
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Dose finding
(single application)
Vaccinees from
Switzerland (Geneva),
Gabon, Kenia
Analysis:
Clinical data
Immunological readout
Our Role (Spiez Lab):
Assessment of neutralization capacities in approx. 150 vaccines, serum
from 2 time points -> 300 serum to test in the BSL-4 Lab
VSV EBOVAC phase I/II dose finding trial
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Establishment of Mobile Laboratories for Pathogens up to Risk Group 4
in Combination with CBRN Capacity Building in Sub-Saharan Africa"
Funded by the European Comission
Build the capacity to bring state-of-the-art technology, diagnostics and highly trained scientists
from Europe and Sub-Saharan African countries to the field in case of infectious disease
epidemics in Europe and Sub-Saharan African countries.
http://www.emlab.eu
• Bernhard-Nocht-Institute for Tropical
Medicine, Hamburg, Germany
• Institute for Lassa Fever Research and
Control (ILFRC), Irrua Specialist Teaching
Hospital, Irrua, Nigeria
• National Institute for Medical Research,
Dar es Salaam, Tanzania
• Bundeswehr Institute of Microbiology,
Munich, Germany
• Istituto Nazionale per le Malattie Infettive
"L. Spallanzani", Rome, Italy
• Institute for Virology, Philipps-University
Marburg, Marburg, Germany
• Laboratoire P4 Inserm Jean Merieux,
Lyon, France
• Spiez Laboratory, Spiez, Switzerland
• Public Health England, Microbiology
Services Division-Colindale, London, UK,
and Microbiology Services-research
Salisbury, UK
Field mission with the European Mobile Laboratory
(EMLab) and MSF in May/June 2014
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
• Modular and rapidly deployable
mobile laboratory in 15 boxes
(checked baggage)
• 3 Lab Units (Tanzania, Nigeria,
München)
• Diagnostic of P3 und P4-
pathogens by PCR, antibody
detection and rapid tests
• Regular trainings und workshops
• Deployable for outbreak
missions
EMLab Concept
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Transfer to Guéckédou
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Hospital tent and medical care workers
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Transfer of Patients to the hospital
Transportation of
infected persons !!
Decontamination of
cars and car
equipment !!
Bad protection
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
MSF protective equipment for patient care
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Sampling by MSF and transfer to the EMlab
MSF Camp (overview)
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
RNA extraction & Mastermix prep PCR amplification (SmartCycler)
Inactivation of patients samples in the glovebox
Sample inactivation by Jasmine Wastebag
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
MSF Logistic
Preparation of the desinfection
solution (Chavel)
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Bundesamt für Bevölkerungsschutz BABS
LABOR SPIEZ
Bild: «biological weapons»
Heinz Sager, 2002
www.labor-spiez.ch
Thank you very much for your attention