animal virology - medilabsecure

Preventing vector borne diseases around the Mediterranean and Black Sea regions

Workpackage 2- Animal VirologyAnimal Health Research Center (INIA-CISA) Madrid, Spain

Miguel Ángel Jiménez ClaveroElisa Pérez RamírezJovita Fernández Pinero

Paris, June 25-27th 2018

MediLabSecure Global Conference

MediLabSecure Global Conference, Institut Pasteur, Paris, June 25-27 2018

Why animal virology?

• Emerging viral zoonoses: growing concern


Why animal virology?

• Aims:

– Creating a network of animal virology labsaround the Mediterranean and Black Sea.

– Increase diagnostic capacity, harmonize methodsand promote preparedness for emerging zoonoticviral diseases in this region.

– Strengthen links with other One Healthcomponents for a better disease surveillance and control.

Animal virology: what for?


One Health in progress!

HUMAN

ENTOMO-LOGICAL

ANIMAL

WP2 Activities

Dissemination

Assessmentof needs

Activities

Workshops

EQA

EQA follow-up meeting

Regional meetings

• SPECIFIC NEEDS

CCHFV pilotstudy

Technical assistance

On-site visits

- Bottom-up strategy- Pathogens of interest.- Separated diagnostics

workshops: - Molecular methods.- Serological methods.

- Wide interest in EQAs.

HEADS OF LABS MEETING – January 2015


ASSESMENT OF NEEDS


MOLECULAR DIAGNOSIS OF RELEVANT ZOONOTIC ARBOVIRUSES

15-19 June 2015 INIA-CISA Madrid (Spain)

• Focus on WNV and RVFV.• Promote transfer of biosafety best laboratory practices• Increase general knowledge on arboviruses in MLS region• Promote networking and collaboration among participants

theoretical sessions and hands-on training in BSL-3 lab

17 participants15 countries15 vet labs + 2 entomology labs

WORKSHOPS

WORKSHOP 1 - Evaluation

• Unique opportunity to visit a BSL3 lab: first experience for 82%.Considered by all participants as a relevant added value.

• Discussions and exchange among vets and entomologists.• Transference of advanced molecular techniques, extremely

appreciated by all the participants. Easy to incorporate to the labs.

POST- WORKSHOP EVALUATION



SEROLOGICAL DIAGNOSIS OF RELEVANT ZOONOTIC ARBOVIRUSES

4-8 April 2016 INIA-CISA Madrid (Spain)

• Focus on WNV and RVFV.• Provide skills and tools to select best serological methods, adapted to

the epidemiological situation and available resources.• Promote transfer of biosafety best laboratory practices.• Increase general knowledge on arboviruses in MLS region.• Promote networking and collaboration among participants

theoretical sessions and hands-on training in BSL-3 labs

16 participants16 countries


• Unique opportunity to visit a BSL3 lab. Considered by all participants as a relevant added value.

• Biosafety sessions (combination of theory + practice).• Virus neutralization training for WNV serological confirmation.• Public presentation of results, discussion and trouble shooting.



INTRODUCTION TO THE MOLECULAR ANALYSIS OF RELEVANT ZOONOTIC PATHOGENS -- BIOINFORMATICS

22-26 May 2017---- INIA, Madrid (Spain)

• Provide bioinformatic skills and tools to perform basic sequence analysis and phylogeny.

• Focus on AIV and WNV.• Promote networking and collaboration among participating

countries.

theoretical sessions and hands-on training



• Quality of the content and the trainers.• Practical exercises with real sequences of pathogens of interest• Personalized assistance during the practical sessions.• Training material (sequences and step-by-step presentations)• Friendly atmosphere: fruitful discussions.



MOLECULAR DIAGNOSIS

(WNV &RVFV)

SEROLOGICAL DIAGNOSIS

(WNV & RVFV) WNV (molecular &

serological)

RVFV(molecular &

serological)

Workshop 1 2015

Workshop 2 2016

EQAs 2017

17 participant labs17 countries

1) Evaluate the laboratory capacity to correctly apply molecular andserological diagnostic techniques for WNV and RVFV detection fromanimal samples.

2) Evaluate the capacity of the laboratories to properly interpret theresults of the various techniques to make a final diagnosis.

Evaluate the degree of learning during the workshops and the capacity of the participants to incorporate the diagnostic techniques into their own

laboratories.


EXTERNAL QUALITY ASSESSMENTS (EQAs)

MOLECULAR PANELS

- 10 samples each panel- Different lineages and strains- Mimic “real” samples (virus

spiked in different organsfrom several species)

SEROLOGICAL PANELS

- 10 samples each panel- Sera from naturally infected

horses (WNV) or vaccinated sheep (RVFV)

POSITIVE CONTROLS

- Extraction and reaction controls

- WNV L1, L2, USUV- RVFV

- RT-PCR reagents- Primers and probes- RNA extraction kit- ELISA kits

REAGENTS AND KITS


Preparation of panels

ShipmentSubmission

of results

Individual reports

Global report

EQA – Results

Molecular panels

Serological panels

94%

6%

Blocking ELISA (16 labs)

100% correct 90% correct

56%

44%

IgM ELISA (16 labs)

100% correct 70-90% correct

WNV

WNV

RVFV

RVFV

The results demonstrated the competence of most of the participant labs (infrastructure and expertise) to face the

molecular and serological diagnosis of WNV and RVFV

Valoración de los participantesEQAs - feedback from participants

The WNV EQA was extremely useful for 94% of the labsThe RVFV EQA was extremely useful for 87.5% of the labs

Before the project: Only 56.3% of labs were able to diagnose WNVOnly 37.5% of labs were able to diagnose RVFV

Incorporation of the diagnostic techniques: 62.5% of labs

The most valuable material:Positive controls > panel (weak positives) > ELISA kits > PCR reagents

MEETINGS


1. INTERSECTORAL MEETINGS (Regional meetings)

- Belgrade, Serbia --- November 2016- Tunis, Tunisia --- July 2017

2. SECTORAL MEETING (Veterinary labs)

- EQA follow-up & scientific technical meeting of theanimal virology network ---- Madrid --- November 2017


EQA FOLLOW-UP & SCIENTIFIC-TECHNICAL MEETING

EQA FOLLOW-UP SCIENTIFIC-TECHNICAL MEETING(Mini symposium on viral zoonoses)

- Global analysis of results.- Share experience among

labs.- Trouble- shooting.- Distribution of the final

report.

- 13 oral presentations of technical/scientific activities bythe network labs.

- Share knowledge onsurveillance and diagnosis of common threats.

- Promote collaboration amongcountries/labs.


21-22th November 2017 INIA-CISA Madrid (Spain)


Feedback from participants

- Solve common mistakes/technical limitations.- Exchange of experience among labs.- Fruitful discussions about technical aspects of the diagnostic procedures and

the interpretation of results.

The meeting was extremely useful for 100% of participants

EQA follow-up session

- Useful information about recent improvement in diagnosis.- Update on current situation of arboviruses in MLS region.- Countries’ priorities and strategies to tackle diseases.- Similar difficulties in surveillance/diagnosis.- Opportunity to present lab results in a small symposium.- New collaborations among labs.

Scientific-technical session


ON-SITE VISITS

Virology lab - IRVT

TUNISIA

Office nationale de sécurité sanitaires des produits alimentaires (ONSSA)

MOROCCO

Institute of Veterinary Medicine of Serbia

SERBIA

Lebanese Agricultural Research Institute (LARI)

LEBANON

Virology lab- Vet Faculty Ankara University

TURKEY

Rep. Vet-Sanitary & Phytosanitary Lab services

Center (SNCO)

ARMENIA

- Good overview of the facilities, research interest and diagnostic procedures

- Meet personally the Head of the lab and laboratory personnel

- Opportunity to present the MLS objectives and activities to all the staff

- Joint visits: Exchange of knowledge, promote intersectoral collaboration

TECHNICAL ASSISTANCE


Provision of biological materials, reagents and protocols for specific needs

Personalized laboratory assistance

- Molecular and serological diagnosis of WNV & AIV- AIV pathotyping and bioinformatics.

Collaborations with I. Pasteur (WP3)

- RVFV EQA for human virology laboratories- Provision of positive controls.- Participation in WP3 workshops: one health vision.

Morocco, Montenegro and Algeria

Morocco, Turkey, Tunisia, Rep. Macedonia, etc.

Need of validated

serologicaldiagnostic tools

for use in animals

CCHF COLLABORATIVE PILOT STUDY


RM Belgrade, 2016: Gaps in animal diagnosis & surveillance tools

CCHF is a threat to the regionCCHF, a tick-borne zoonosis, is a growing concern

Enrolling animal virology labsconcerned

Search of available tools

Validation study

From: Spengler, JR. Et al Antiviral Res 135:31-47 (2016)From: who.int

1) Screening test

2) Confirmatory test


DISSEMINATION

Thank you very

much for your

collaboration and

we wish to

continue working

together

You guys did a really perfect job from the beginning of the project. Having chance to share the friendly atmosphere with you and other participants is always great chance for me. Thank you for all your efforts and friendship

FEEDBACK FROM BENEFICIARIES

The team at CISA-

INIA did an

incredible job.

You are very friendly.

With you it is very

easy to solve any

question related to

protocols, diagnostic

materials, contacts,

etc.

Good job!

NEXT CHALLENGES

WHAT WE HAVE ACHIEVED…Create collaboration

Generate trustIncrease lab capacities

Incorporate One Health

WHAT WE NEED…Consolidate the network

Get prepared for new threatsFace global changes

Reinforce One Health

ACKNOWLEDGEMENTS


Aykut Ozkul, Ankara University, TurkeyFederica Monaco, IZSAM, ItalyGian Mario Cossedu, IZSAM, ItalyJordi Figuerola, EBD-CSIC, SpainAlejandro Brun, INIA-CISAGonzalo Pascual, INIA-CISARicardo Ramos, Parque Científico, SpainAna Moreno, IZSLER, ItalyFerrán Palero, CEAB-CSIC, SpainIván Sanz, National Influenza Center, Spain

THANK YOU

animal virology - medilabsecure

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