Filoviruses

Dr. Olga Dolnik Philipps-University Marburg

Institute of Virology 24.2.2015

ECDC Training Course – Ebola and Marburg Diagnostic 24th February to 27th February 2015

Robert Koch Institute, Berlin

Taxonomy

Order

Family

Genus

Species

Mononegavirales (ss –RNA)

Rhabdoviridae Paramyxoviridae Filoviridae Bornaviridae

Ebolavirus Marburgvirus Cuevavirus

Zaire

EBOV

Sudan

SUDV

Tai Forest

TAFV

Reston

RESTV

Bundibugyo

BDBV

First documented Filovirus Outbreak

• Marburg, Frankfurt, Belgrade. • Primary cases: Lab workers

with contact to tissue of imported monkeys (Uganda).

• Secondary cases: relatives, health care workers.

• 32 cases, CFR: 25%.

Werner Slenczka Rudolf Siegert 100 nm

Conjunctivitis Enanthema

Thrombocytopenia Diarrhea Exanthema Hepatitis

Hemorrhages

Marburg virus hemorrhagic fever

Imported Marburg fever (Netherlands 2008)

• July 5: – Dutch tourist (41) returning from holiday in Uganda (June 5 – 28) – Presents to the general practitioner with high fever (39°C, 3d) – Admission to the local hospital – Placed in a room with 3 other patients – Malaria diagnostics negative – Bacteriological tests, treatment with ceftriaxone (2 g/day)

• July 7: – Rapid deterioration, liver failure: Hemorrhagic fever as differential

diagnosis included – Transfer to Leiden University Hospital; patient developed rash,

conjunctivitis, diarrhea, liver and kidney failure, hemorrhages. – Extensive bacteriological and virological analyses

• July 10: – Tested positive for Marburg virus at the Bernhard-Nocht-Institut in

Hamburg.

• July 1: – Patient died of cerebral edema.

The Python cave, Uganda

© René Gottschalk © René Gottschalk

Marburg virus RNA in animal species collected in the Goroumbwa gold mine, Durba

Swanepoel et al., EID, 2007

Durba: Bat and human MARV isolates

Phylogenetic tree

Swanepoel et al., EID, 2007

Since 1976 emerging EBOV outbreaks in central Africa

EBOV outbreaks in humans

Ebola virus Reston circulates in Philippines

- Epizootics since 1989 - non pathogenic in humans - pathogenic for cynomolgus macaques (systemic) - symptomatic disease in pigs (lung), asymptomatic infections - transmission to humans (IgG) - REBOV-specific IgG in Rousetttus amplexicaudatus bats

Transmission and Reservoir

Detection of specific antibodies Isolation of virus specific RNA

Distribution of fruit bats in equatorial Africa

Baize et al., 2014

Emergence of Zaire Ebola virus in Guinea

Regional distribution of confirmed and suspected cases of EVD in October 2014 (WHO)

1. Densely populated regions with big cities, Conakry (Guinea), Monrovia (Liberia) und Freetown (Sierra Leone)

2. Poor medical infrastructure in rural regions, traditional burial practices 3. Cross-boarder trade and social activities

Three major factors driving continuous transmission of EBOV in West Africa

Human to human transmission

17

- direct body contact with symptomatic patients - direct contact with body fluids of patients (blood, urine, stool, sweat, saliva, sputum,

semen, breast milk) - direct contact with contaminated surfaces - no air transmission - low kontagiosity (R0 Ebola 1-4, R0 measles 12-18) - moderate thermostability (4°C weeks, room temperature days, 60°C 30min, 5min boiling) - sensitive to UV, detergents, formaldehyde, acids - anti-viral disinfectant (enveloped viruses!), chloric bleach

SW Mitchell end JB McCormic, 1984 Piercy et al., 2010

Prevention

Ebolavirus

19

Glycoprotein, GP

ribonucloproteincomplex

matrix, VP40

lipidenvelope

L. Kolesnikova

1977 W. Slenczka

3´ Leader Trailer 5´ NP 35 40 GP 24 30 L

sGP

Replication cycle

Dolnik et al., Cell Mol Life Sci. 2008

NPC1 actin

Pathogenesis

Nature reviews Immunology 2007

• Incubation time 2-21 days

Symptoms of Ebola virus disease (EVD)

• Beginning with high fever (>38,5°C), muscle pain, nausea

• Multiplication of the virus in vital organs like liver, kidney, spleen and lung which causes organ failure

• abdominal pain, diarrhea

• destruction of the vascular endothelium

• sever cases die due to multi-organ failure and shock

Case definition EVD

clinical, epidemiological, high-risk exposer and laboratory criteria

clinical criteria: fever ≥ 38,6°C and any of the following: - severe headache - vomiting, diarrhea, abdominal pain - unexpected hemorrhagic signs - multi-organ failure epidemiological criteria: within 21 days before the onset of symptoms:- having been in an area with community transmission of EVD - having contact with probable or confirmed EVD case high-risk exposer criteria: - close contact (within 1m) without appropriate personal protection with a probable or confirmed case who was coughing, vomiting bleeding, had diarrhea, unprotected sexual contact with a case up to three month after recovery - direct contact with body fluids from a probable or confirmed case - percutaneous injury or mucosal exposure to body fluids, tissue or laboratory specimens of a probable or

confirmed case - participation in funeral rites with direct exposure to human remains in or from affected area without

personal protection - direct contact with bats, rodents, primates, living or dead, in or from affected area, or bush meat

laboratory criteria: detection of nucleic acid, Ag-ELISA, EM in clinical specimens

Management of patients with haemorrhagic fevers

• Isolation units, Personnel under full protection • Meldung an das jeweils zuständige

Gesundheitsamt.

(Meldepflicht bei Auftreten und beim Verdacht auf hämorrhagisches Fieber (IfSG).

25

BSL 4 Laboratory at the Philipps-University in Marburg

BSL4 Lab

Diagnostic Direct virus detection: Real-time-PCR (Altona Filovirus screen) > RNA detection! Antigen-ELISA (Matrix protein specific antibodies) (Senova) Electron microscopy > detection of virus particles in patient samples Virus cell culture (BSL-4) > CPE

Diagnostic Antibody detection (Serology): IgM-IgG ELISA Indirect Immunofluorescence Test (IFT) Virus neutralization test

Vaccine

Experimental drugs and vaccines in clinical trials

Thank you for your attention!

Questions?