EURL rabies
Collection of samples, techniques, validation and interpretation of the diagnostic methods for the purposes of rabies surveillance are presented here.
Diagnostic methods considered compliant are presented in the WOAH rabies manual. A list of procedures inspired by the WOAH official methods is also proposed.
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Work on rabies by Nancy laboratory for rabies and wildlife
Last update: 23 April 2021
Organising, coordinating, implementing and monitoring oral fox vaccination campaigns against rabies
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In France, in 1968 following the first vulpine rabies cases in the North-East of country, the front advanced rapidly westwards and southwards. In 1971, nine departments, totalling an area of 25000 km² were infected. The laboratory was built in 1971 in the infected area to conduct rabies diagnosis on suspect and found dead animals.
In the 1970s, the laboratory extensively participated in scientific research work for developing oral vaccine baits to control the disease in fox population.
A number of experimental trials were undertaken to check efficacy and safety of vaccines before their marketing, in collaboration with other international groups involved in rabies activities. In 1986, the laboratory tested the first available methods of oral vaccination with baits (chicken head baits) deposited in the field by manual distribution. From 1990, the overall organisation of the oral vaccination campaigns has been centralised: the Anses-Nancy Laboratory for rabies and wildlife was requested by the national competent authority to organize and plan the oral vaccination of foxes and, in close cooperation with its delegates, to implement and evaluate the programmes.
From 1988 to 1998, year of the last recorded case of vulpine rabies in France, the laboratory planned the strategy and implemented oral vaccination campaigns in infected areas twice a year using helicopters. These campaigns were then prolonged until 2003 along Northern and Eastern borders and were performed once again in 2005.
Several field trials have been conducted to improve the efficiency of oral vaccination programme, particularly to promote cubs' coming into contact with baits and also to counterbalance the increase of the fox population.
Rabies surveillance and vaccination monitoring
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Based on the collection of animals found dead, road killed, suspect animals as well as all animals having contamined humans, the national surveillance network is organised as represented below :
- In case of human contamination (bite, scratch), animal cadavers are sent by the veterinary services to the Pasteur Institute.
- In all other cases, they are submitted by the veterinary services to Anses for rabies testing.
Rabies surveillance is the key element of rabies control and prevention programmes. It is based on laboratory investigations on samples taken from susceptible suspect animals to collect sufficient spatial and temporal descriptive data used to build the control programme.
The national network for animal rabies surveillance is centralised at Anses-Nancy laboratory for rabies and wildlife. The incidence of rabies in animals is currently assessed throughout all the country by undertaking diagnosis on suspect animals using the OIE and WHO reference tests, i.e. the fluorescent antibody test and the real time RT-PCR (WHO, 2018; OIE, 2018). The positive cases are systematically investigated by sequencing to characterise the virus strains of isolated lyssaviruses, and also to study the circulating field isolates in the country.
The monitoring of the efficacy of oral vaccination programmes was conducted in vaccinated areas by:..
- Assessing bait uptake by testing the target species (foxes) in vaccinated areas for biomarker (tetracycline) occurrence and assessing age categories of foxes marked by tetracycline.
- Assessing rabies antibodies in target species in vaccinated areas: a cell virus neutralisaion method by using an ELISA previously validated for wildlife sample titration.
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Technical competences in rabies
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The laboratory currently uses the folowing laboratory methods (Table 1):
Table 1 | ||
Areas | Methods | Comments |
Diagnostic |
FAT RTCIT DRIT Conventional RT-PCR SYBR Green RT-PCR TaqMan RT-PCR |
|
Serology |
FAVN test Indirect ELISA Competitive ELSA |
Used on animal sera and animal immunoglobulins |
Vaccine potency testing |
NIH Pharmacopeia test TSA |
|
Monitoring of vaccination programmes |
Biomarker determination Age determination |
Both on tooth samples |
Virus titration |
On cells On animals |
The laboratory is experienced in the development, validation and standardisation of laboratory methods, mainly for diagnosis and serology, and regularly organises international workshops, inter-laboratory studies and proficiency tests.
The laboratory is enrolled in quality assurance (Table 2):
Table 2 | ||
Standard |
Test | Accreditation body |
NF EN ISO/IEC 17043 | Organization of rabies serological proficiency test |
COFRAC (since May 2017) |
NF EN ISO/IEC 17025 | FAT (Lyssavirus antigen) and RTCIT (Lyssavirus) |
COFRAC (since October 2012) |
NF EN ISO/IEC 17025 | Molecular Biology (Lysaviruses RNA detection) |
COFRAC (since March 2020) |
NF EN ISO/IEC 17025 | Control of efficacy of live oral vaccines in wildlife |
Mutual Joint Audit (EDQM) (since January 2012) |
NF EN ISO/IEC 17025 | Control of efficacy of inactivated rabies vaccines for veterinary and human use (challenge test and serological test) |
Mutual Joint Audit (EDQM) (since March 2008) |
NF EN ISO/IEC 17025 | Rabies virus seroneutralisation test (FAVN test) |
COFRAC (since February 2008) |
The laboratory developped several methods (Table 3):
Table 3 | ||
Name of the test developed | Use | References |
Straw sampling kit | Simplified technique dor the collection, storage and shipment of brain material from animals for rabies diagnosis |
Barrat et al., 1986 - WHO, 1987 - WHO, 1996 - WHO, 2018 |
Simplified test for potency testing of inactivated veterinary vaccines | Potency testing of vacines using one dilution of the vaccine |
Aubert, 1982 - WHO, 1996 - European Pharmacopeia, 2014 |
FAVN (Fluorescent Antibody Virus Neutralization) test | Titration of seroneutralising rabies antibodies in human and animals blood samples |
Cliquet et al., 1998 - WHO, 2018 - OIE, 2018 |
Indirect ELISA | Titration of rabies virus antibodies in wildlife samples for vaccination monitoring | Cliquet et al., 2000 |
Paper-based diagnosis for sample sending through normal postal means | Impregnation of rabies suspect brain tissue to allow safe and cost effective shipment of diagnosis samples | Picard Meyer et al., 2007 |
Paper-based technology to collect blood samples of wildlife | Impregnation of blood samples to allow simple collection and storage of field samples for further serological testing using ELISA | Wasniewski et al., 2014 |
Hemi-nested RT-PCR method | Specific determination of EBLV-1 Lyssavirus | Picard-Meyer et al., 2004 |
The laboratory is also organising proficiency tests (Table 4):
Table 4 | ||
Proficiency test | Since | Frequency |
Dog and cat rabies serology | 2000 | Annual since 2009 |
Rabies diagnosis (FAT, RTCIT, RT-PCR, Real Time RT-PCR) | 2002 |
Annual since 2009 Every two years since 2017 |
Tetracycline biomarker and age determination | 2010 | Every two years |
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Research activities on rabies
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A the present time, the research work of the laboratory is mainly intended to increase knowledge on the pathogenicity of certain rabies viruses (those that infect European bats), to develop experimental models and to investigate the transmission of bat lyssaviruses to other bats and to non flying animals.
0 November 2014
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FRENCH AGENCY FOR FOOD, ENVIRONMENTAL AND OCCUPATIONAL HEALTH & SAFETY
Nancy Laboratory for Rabies and Wildlife
Batiment H
Technopôle Agricole et Vétérinaire
CS 40 009
54220 Malzéville - France
GPS localisation: 48.72635718594794, 6.181035981226453
Annual review of analysis performed in NRLs
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Rabies disease
Last update: 15 January 2020
Rabies is a fatal zoonotic disease (animal disease that can be transmitted to humans) caused by a virus of the genus Lyssavirus. Excreted in the saliva of infected mammals in the final phases of the disease, the virus is generally transmitted to another animal or to humans through biting. Contamination may also occur if the saliva of an infected animal comes into contact with an open wound or a mucous membrane. Without post-exposure treatment prior to the onset of clinical signs, the disease is invariably fatal.
Rabies, which causes over 59,000 human deaths a year worldwide, is found all over the world, except in certain areas such as Antarctica. Several European countries have become rabies-free in non flying mammals thanks to oral vaccination programmes of wildlife.
There are 16 different rabies virus species, seven of which transmission to human has already been notified. Those species are mainly differentiated according to the animal host species. Rabies due to rabies virus species (RABV) is responsible for most human and animal rabies cases.
In industrialised countries, rabies persists mainly in wild animals, whereas in many developing countries it remains an endemic disease, with the domestic dog as principal reservoir and main source of human contamination.
In European countries, rabies in dogs was eliminated several decades ago, but it continued to persist and spread in fox and racoon dog populations. Thanks to oral vaccination campaigns conducted in wildlife, the incidence of rabies in both domestic and wild animals in the EU has drastically reduced. Rabies has been eliminated from the Netherlands, Finland, Switzerland, France, Belgium, Luxembourg, Czech Republic, Austria, Germany, Estonia, Italy, Latvia and Slovenia.The elimination of rabies in non-flying mamals (RABV) in the European Union will be reachable in the next years. In 2018 and 2019, eight (from 3 EU countries) and five (from 2 EU countries) cases, respectively, were reported in the EU.
To detect timely any suspect animal, the rabies situation in all Europe should be continuously monitored, based on surveillance programmes. The illegal importation of infected cats and dogs from endemic countries remains a major concern, with regular rabies alerts occuring (ProMED). In Europe, bat rabies cases are attributed to five different lyssavirus species. While European bat lyssavirus types 1 and 2 are responsible for most bat rabies cases, Bokeloh bat lyssavirus, West Caucasian bat lyssavirus and Lleida bat lyssavirus have occasionnaly been isolated.
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