LRUE Fièvre Aphteuse

URL: 
https://eurl-fmd.anses.fr
Background type: 
color
Intro [EN]: 

The European Union Reference Laboratory organises Proficiency Tests (PTs) on detection and typing of Foot-and-Mouth disease virus in accordance with Council Directive 2003/85/EC and Regulation (EU) No 2017/625.

PT panels for FMDV and SVDV detection are established for virus isolation, RT-PCR, Antigen-ELISA and VP1 sequencing. PT panels for FMD antibody detection are established for FMDV NSP antibodies, FMDV SP antibodies and SVDV antibodies detection by ELISAs or/and virus neutralisation test.

The proficiency test (PT) for FMD virus and SVD virus and antibody detection is organised annually. All EU national reference laboratories (NRLs) and other official laboratories of the network are invited to participate.

Intro [EN]: 

The European Union Reference Laboratory yearly organizes a workshop, especially intended for the EU national reference laboratories (NRLs) and other official laboratories of the network.

The purpose of these workshops is to inform the participants about the activities of the EURL, to present and discuss the results of the PT, to present and discuss scientific research projects and to exchange information on EU legislation, methods and other relevant subjects.

 Registration to the next workshop

Intro [EN]: 

The EURL for FMD, among other missions, gathers data and information on the methods of diagnosis and differential diagnosis used in the different National Laboratories.

A list of methods inspired by the OIE official methods is available on this website.

Intro [EN]: 

The EURL for FMD supports the functions of National Laboratories by storing and supplying reagents and materials for use in diagnosis of foot-and-mouth disease such as virus and/or inactivated antigens, standardised sera, cell lines and other reference reagents.

A list of reagents and material is available on this website.

Display Name: 
Foot-and-Mouth disease
Title [FR]: 
Méthodes
Title [EN]: 
Methods
Title [FR]: 
Réactifs disponibles et matériaux de référence
Title [EN]: 
Available reagents and reference materials
Methods Background: 
Materiau Background: 
Intro [EN]: 

The European Union Reference Laboratory organizes each year trainings specifically intended for the EU national reference laboratories (NRLs) and other official laboratories of the network.

The purpose of these sessions is to implement the further training of experts in laboratory diagnosis with the goal to harmonize the diagnostic techniques at the European and International levels.

Registration to the next training

Samples treatment

 

Some samples can be used directly, without any preparation. Epithelial flaps have to be homogenised and clarified. Find this document here.

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Duplex one-step real-time RT-PCR assays

 

After RNA extraction, the test first relies on the reverse transcription (RT) of the RNA of FMD virus into complementary DNA (cDNA). The cDNA then undergoes PCR amplification by a DNA polymerase that uses primers and a TaqMan® probe, labelled with a FAM or VIC fluorochrome at the 5' end. The two enzymatic reactions are performed successively in the same tube (one-step, real-time RT-PCR).

Due to the high level of mutations generated during the replication of RNA viruses, it is recommended to use two specific primer pairs in parallel targeting two different regions of the FMD genome: the IRES region and the encoding region for the RNA-dependent RNA polymerase 3D.

In addition, a third pair of primers is used to detect the endogenous ß-actin cellular gene (present in the extracted cellular RNA fraction). Detection of this gene acts as a control for the extraction step and absence of PCR inhibitors.

This is a duplex reaction. In a single reaction, two targets are detected simultaneously: either the IRES + ß-actin targets, or the 3D + ß-actin targets. The probe for the FMD target is labelled with a FAM fluorochrome and the probe for the ß-actin target with a VIC fluorochrome.

Find this document here.

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Republic of Moldova

Ministry of Agriculture and Food Industry: Republican Veterinary Diagnostic Centre

English

Belarus

Belarusian State Veterinary Centre

English

Ukraine

State Research Institute of Laboratory Diagnostic and Veterinary Sanitary Expertise

English

Bosnia - Herzegovina

Veterinary Institute "Dr. Vaso Butozan"

English

Switzerland

Institute of Virology and Immunology

English

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