FMD Reference Laboratory

WRLFMD Team: Valerie Mioulet, Nick Knowles, Anna Ludi, Ginette Wilsden, Kasia Bachanek-Bankowska, Lissie Hendry, Jemma Wadsworth, Britta Wood, Barsha Thapa,

Bob Statham, Abid Bin-Tarif, Ashley Gray, Clare Browning, Beth Johns, Mark Henstock, Alison Morris, David Paton, Veronica Fowler, Nick Lyons, Dexter Wiseman,

Julie Maryan, Sarah Belgrave

FMD in West Africa:What we know and what we don’t know

Donald King

World Reference Laboratory for FMD (WRLFMD)donald.king@pirbright.ac.uk

• Containment Level 2 Laboratory• Commissioned 2016

• High-containment Level 3+ Laboratory

• Commissioned 2015• “Not-for profit” research

institute • UK and International Reference

Centre for livestock viral diseases (including: FMD, BT, PPR, ASF, AHS, Capripox, Marek’s Disease)

• Work on behalf of OIE, FAO, EuFMD and EU

The Pirbright Institute

Global FMD NetworksOIE/FAO FMD Laboratory Network

• Exchange of data between OIE and FAO Reference Centres

• Collection and testing of samples from countries where FMD is endemic

• Global surveillance and changing threats

• Harmonised and improved lab capacity

Brussels, Belgium – November 2015

FMD: Conjectured global status in 2016• Seven FMDV serotypes• Seven endemic pools requiring tailored diagnostics and vaccines• Global distribution of serotypes: O>A>SAT 2>SAT 1> Asia 1• Serotype C not detected since 2004• Last clinical cases in South America in 2012

• West Africa considered to be endemic for FMD• Serotypes O, A and SAT 2 (SAT 1?)

Importance of sampling FMD outbreaksOIE/FAO FMD Lab Network Data for 2015

*

• Baseline information that contributes to the understanding of the impact and burden of disease

• Provides sequence data to understand the trans-boundary movements of FMD virus

• Vaccine strain selection and recommendations

• Globally:

1034 (2013), 3245 (2014), 2079 (2015)

• West Africa samples*:

40 (2013), 178 (2014), 147 (2015)

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1000 km

600 mi

KenyaCongoGabon

Cameroon

Central

African

Republic

South

Sudan

Ethiopia

Somalia

Sudan

Egypt

Nigeria

Togo

GhanaCôte

d’IvoireSierra

Leone

Guinea

Mali

Burkina

Faso

Guinea-

Bissau

Senegal

Mauritania

Morocco

Algeria

Niger Chad

Libya

Liberia

Uganda

TanzaniaBurundi

Democratic

Republic of

the Congo

FMD reported in the last 10 yearsFMD not confirmed in the last 10 yearsUnknown FMD status (never confirmed)

19742005

19961996

20021973

1999

1999

A, 2006

SAT2, 2014O, 2006

A, 2006 O, 2015

SAT2, 2005

O, 2006

SAT2, 2009

O, 2010

A, 2010Benin

O, 2014

A, 2013

SAT2, 2012

O, 2005

A, 2013

SAT2, 2013

Eritrea

Tunisia

20042003

SAT 1 has not been confirmed in West Africa since 1981

Does the current level of sampling of field cases provide an accurate picture of the true extent of FMD in the region?

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1000 km

600 mi

KenyaCongo

Gabon

Cameroon

Central

African

Republic

South

Sudan

Ethiopia

Somalia

Sudan

Egypt

Nigeria

Togo

GhanaCôte

d’IvoireSierra

Leone

Guinea

Mali

Burkina

FasoGuinea-

Bissau

Senegal

Mauritania

Morocco

Algeria

Niger

Chad

Libya

Liberia

Uganda

TanzaniaBurundi

Democratic

Republic of

the Congo

Ind-2001dInd-2001d Ind-2001d

Ind-2001d

EA4

EA3

EA2

EA4

EA3EA3

EA3

EA3

EA2

EA2

EA2

EA3

WAWAWA

WA

WA

PanAsia-2

WA

WA

FMDV O

EA-3

PanAsia-2

EA3

Serotype O lineages

O/WA Maintained within West Africa

O/EA-3 Multiple introductions into the region

FROMASIA

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KenyaCongo

Gabon

Cameroon

Central

African

Republic

South

Sudan

Ethiopia

Somalia

Sudan

Egypt

Nigeria

Togo

GhanaCôte

d’IvoireSierra

Leone

Guinea

Mali

Burkina

FasoGuinea-

Bissau

Senegal

Mauritania

Morocco

Algeria

Niger

Chad

Libya

Liberia

Uganda

TanzaniaBurundi

Democratic

Republic of

the Congo

FMDV A

G-VI

G-VI

G-I

G-I

G-VII

Iran-05

G-IV

Iran-05

G-IV

G-VII

G-VII

G-IVG-IV

G-VI

G-IV

G-I

G-IV

G-IV

A/AFRICA/G-IV Maintained in West Africa

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600 mi

KenyaCongo

Gabon

Cameroon

Central

African

Republic

South

Sudan

Ethiopia

Somalia

Sudan

Egypt

Nigeria

Togo

GhanaCôte

d’IvoireSierra

Leone

Guinea

Mali

Burkina

FasoGuinea-

Bissau

Senegal

Mauritania

Morocco

Algeria

Niger

Chad

Libya

Liberia

Uganda

TanzaniaBurundi

Democratic

Republic of

the Congo

FMDV SAT 2

VII

VII

VIIVII

VIIXIII

XIIIVII

IV

IV

VII

2014: SAT 2/VII outbreaks in Mauritania

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KenyaCongo

Gabon

Cameroon

Central

African

Republic

South

Sudan

Ethiopia

Somalia

Sudan

Egypt

Nigeria

Togo

GhanaCôte

d’IvoireSierra

Leone

Guinea

Mali

Burkina

FasoGuinea-

Bissau

Senegal

Mauritania

Morocco

Algeria

Niger

Chad

Libya

Liberia

Uganda

TanzaniaBurundi

Democratic

Republic of

the Congo

FMDV SAT 1

I (widespread)

I (widespread)

IX (very limited in 2007)

??? SAT 1 ???

1975-81 SAT 1 cases reviewed by Sangare et al., 2003 1999-2003:Wildlife serosurvey – Di Nardo et al., 2015No recent field isolates or sequences from West Africa

• FMD is a trans-boundary disease

o Factors underpinning the epidemiology of FMD in West Africa are not well understood

o Which FMD serotypes/topotype circulate in each of the countries?

o Economic impact of FMD

• Research gaps?

o Understanding the challenges: Knowledge about how each of the FMD viral lineages are maintained in the region or are introduced from countries outside of the region

o Tools for FMD Control: Robust evidence for the selection and use of vaccines in the region: are effective vaccine available for the strains that circulate in the region?

o Response: Tools for the rapid detection and characterization of FMD viruses causing field outbreaks

• Importance of the sample collection from FMD outbreaks in the field

o To feed real-time lab data back to FMD control programmes

• WRLFMD and the OIE/FAO Lab Network welcome sample submissions from member states

• Testing is free of charge

• Support from EuFMD for sample shipments

• Contact: donald.king@pirbright.ac.uk

Summary and (obvious?) laboratory priorities

Acknowledgements• Support for the WRLFMD and

research projects

• Collaborating FMD Reference Laboratories and field teams

• Partners within the OIE/FAO FMD Lab Network

Reports:

www.wrlfmd.org

• FMD is endemic in the East Africa region

• Four FMDV serotypes have been recently reported in the country (A, O, SAT1, SAT2)

• Factors associated with outbreaks are not clearly known

• Links to outbreaks in neighbouring countries

• Epidemiology of FMD is complicated by involvement of wildlife (Buffalo)

Case Study: Tanzania

NEG POS

• LAMP products can be detected using LFD

-FITC and Biotin labelled oligos

-Multiplexing is possible

• Amplification can be performed using a water bath

o Simple

o Rapid

o Inexpensive?

o Basis of a disposable test?

• Could be deployed into multiple locations in the event of FMD outbreaks

Waters et al., 2014: PLoS ONE

RT-LAMP: simple formats/minimal equipment

Pirbright Team in Tanzania: 2014 and 2015

Emma Howson, Bryony Armson and Veronica Fowler

KasiaBankowska

• Field trial in Tanzania

• Pilot data for FMDV

• Equivalent performance to lab-based real-time RT-PCR

• Evaluation of new dry-down reagents and enzymes

RT-LAMP in the field:

Paulo-Fupi Raphael in Tanzania (2013)

• FMDV can be detected by real-time RT-PCR using nucleic acid recovered from positive LFDs

• RNA is stable > 1 month

• Possible to generate larger RT-PCR amplicons

o Strain characterisation (VP1 –full genome)

o “live” virus recovery by electroporation into BHK cells

• Use of LFDs to ship FMDV positive material to Ref. Labs?

Fowler et al., 2014: PLoS ONE

Not only local testingWhat else can we use Ag-LFDs for?