Ebola outbreak - · PDF file• The Ebola Virus • Transmission ... Mission of the biology branche (1995 ... Project Biocontainment Laboratory

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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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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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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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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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LABOR SPIEZ

The Ebola Virus

Replikation cycle:

May infect and propagate in most of the human tissue

100 nm

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LABOR SPIEZ

The Ebola Virus


• Variable „human relevant“ spezies (Ebola, Marburg)


• 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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LABOR SPIEZ

The Ebola Virus

• 2007

Uganda

• 37 deaths

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LABOR SPIEZ

The Ebola Virus


• Variable relevant spezies (Ebola, Marburg, Reston)


• 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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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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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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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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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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Transmission (Cycle)

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Ebola outbreak in Africa

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Distribution in 2014

June July August


The indicator case was already registered in September

2013 but not recognized as an Ebola infection.

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Situation as of February 2015

Source: CDC, Atlanta, USA

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Situation ausserhalb Westafrikas

6.Oktober 2014

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Total cases February 2015

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Guinea (66.7% )

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Sierra Leone (29%)

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Liberia (42%)

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First isolates from Ebolastrain Zaire (Guinea)

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Marburg outbreak

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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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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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Vaccine trials 1

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Vaccine Trials 2

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LABOR SPIEZ

Tasks of SPIEZ LABORATORY

Biocontainment Laboratory

BSL-4

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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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Birds view

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LABOR SPIEZ

Organigram

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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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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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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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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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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


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Containment Concept

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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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Biocontainment Laboratory: Design

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The lab staff is protected from the pathogens


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Society / environment is protected from pathogens

Filtration of incomig and outgoing air

using HEPA filters (H14)

Waste water treatment (sterilisation)


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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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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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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

http://www.google.ch/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&uact=8&ved=0CAcQjRw&url=http://viralzone.expasy.org/all_by_species/21.html&ei=tRzGVJPKBoXAPIOugJAN&bvm=bv.84349003,d.bGQ&psig=AFQjCNHjrcOORhZiH-NXhlSLWTrKGZvZPA&ust=1422355993981649

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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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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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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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Transfer to Guéckédou

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Hospital tent and medical care workers

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Transfer of Patients to the hospital

Transportation of

infected persons !!

Decontamination of

cars and car

equipment !!

Bad protection

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MSF protective equipment for patient care

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Sampling by MSF and transfer to the EMlab

MSF Camp (overview)

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RNA extraction & Mastermix prep PCR amplification (SmartCycler)

Inactivation of patients samples in the glovebox

Sample inactivation by Jasmine Wastebag

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MSF Logistic

Preparation of the desinfection

solution (Chavel)

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LABOR SPIEZ

Bild: «biological weapons»

Heinz Sager, 2002

www.labor-spiez.ch

[email protected]

Thank you very much for your attention

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Ebola outbreak - · PDF file• The Ebola Virus • Transmission ... Mission of the biology branche (1995 ... Project Biocontainment Laboratory