Rinderpest and Peste des petits ruminants
OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / Rinderpest and Peste des petits ruminantsAddress of laboratory: / CIRAD-BIOS, Control of Exotic and Emerging Animal Diseases
Campus International de Baillarguet TA A-15/G, 34398 Montpellier Cedex 5, France
Tel.: / (+33[0]4)67.59.37.98
Fax: / (+33[0]4) 67.59.38.50
e-mail address: /
website: /
Name of Head of Laboratory (Responsible Official): / Dr Geneviève Libeau
Name of OIE Reference Expert: / Dr Geneviève Libeau
Name of writer of this report
(if different from above): / Dr Geneviève Libeau
Summary of general activities related to the disease
1.Test(s) in use/or available for the specified disease at your laboratory
List of tests available for the diseases.
Test / For / Specificity / Surveillance / Research / TotalELISA / Antibody / RP-specific
PPR-specific / 532
532 / 500
500 / 1032
1054
ICE-ELISA / Antigen / RP-specific
PPR-specific / 0
0 / 100
100 / 100
100
VNT / Antibody / RP-specific
PPR-specific / 0
0 / 0
0 / 0
0
Virus Titration
/ Virus / RP-specificPPR-specific / 0
2 / 1
100 / 1
100
RT-PCR
/ Genome / RP-specificPPR-specific
Pan-morbillivirus
DVM / 261
261
261
0 / 100
100
0
0 / 361
361
261
0
QRT-PCR
/ Genome / RP-specificPPR-specific
Pan-morbillivirus / 0
100
0 / 0
600
0 / 0
700
0
Peptide ELISA
/ Antibody / PPR-specific / 0 / 0 / 0Sequencing
/ Genome / RPPPR
DMV / 0
30
0 / 1
10
0 / 1
40
Cell culture
/ Virus isolation / RPPPR
DMV / 0
3
0 / 0
5
0 / 0
8
0
The competitive ELISA test (C-ELISA) for peste des petits ruminants (PPR) antibody detection is based on the anti- nucleoprotein (N) monoclonal antibody (Mab). Diagnosis is completed by the use of the C-ELISA based on a specific anti-haemagglutinin (H) antibody. We follow the same approach for rinderpest (RP) by using two C-ELISA based on specific anti-H and anti-N. The neutralisation test is also implemented on sera detected positive by ELISA. Although it is time consuming, the protocol based on different tests for differential diagnosis is necessary, as not a single test is actually very specific.
Serosurveillance is targeted on domestic ruminants and wildlife. The diagnostic activity is carried out in support to countries experiencing PPR or country willing to monitor the rinderpest situation in order to meet the requirements contained in the OIE pathway and Chapter 1.1.6 ( of terrestrial Animal Health Code.
2.Production and distribution of diagnostic reagents
The immunocapture (ICE) ELISA test for the differential diagnosis of RPV and PPRV and the N-PPR competitive ELISA are prepared by CIRAD and supplied through a private company. One kit is prepared and supplied by CIRAD: the N-RP competitive ELISA.
Diagnostic reagents and ELISA kits for the detection of antigen and antibodies were supplied to the following countries: UK, Bangladesh, Sudan, Israel, Ethiopia, Burkina, Mali, Turkey, Morocco, etc...
Number, type of kits, vaccine, strains and of reagents:
- Immunocapture ELISA: 16
- C-ELISA NRP: 0
- C-ELISA NPPR: 55
Type of reagents:
- RNA, cDNA,
- Wild PPR strains, live or inactivated
- Mabs, reference sera, cell lines,
- Primers,
- Plasmids containing PPR genes.
Activities specifically related to the mandate
of OIE Reference Laboratories
3.International harmonisation and standardisation of methods for diagnostic testing or the production and testing of vaccines
With partners from Europe and Asia, the reference laboratory is involved in the EU project named Epizone (Network of Excellence for Epizootic Disease Diagnosis and Control) which objective is the exchange of information and the sharing of knowledge with impact on (a) a common approach to control animal diseases, (b) a better understanding of the distribution of epizootic diseases present in Europe and (c), an enhanced capacity to give advises to governmental entities, NGO and project holders.
In addition, to enforce expertise of other laboratories from developping countries in PPR diagnosis, inter-laboratory ring trials for PPR antibody detection wereorganized. Reference materials and protocols were prepared and shared among the different participants.
We are also involved in a Coordinated Research Project with the FAO/ IAEA Centre for ELISA and Molecular Techniques in Animal Disease Diagnosis. The project is entitled Early and Sensitive Diagnosis of Peste des Petits Ruminants. The main objectives of this project are to standardize and validate new molecular-based (real time PCR, LAMP) PPR diagnostic tools and penside tests for early and sensitive diagnosis of PPR.
4.Preparation and supply of international reference standards for diagnostic tests or vaccines
International standards are widely distributed as well as diagnostic kits. For vaccine technology transfer in other countries, the master seed of the PPR 75-1 vaccine is made available and the production technology is implemented. Once implemented in a country, the Reference Laboratory, as part of the quality control of the product, also follows up the quality and safety of the vaccine production. In 2010, a request for vaccine quality control was satisfied.
5.Research and development of new procedures for diagnosis and control
In 2010 research projects were followed in order (1) improve our approach to control the disease by the generation of biological antiviral, (2) improve diagnostic tools by the development of quantitative RT-PCR for PPRV, (3) improve our knowledge of PPRV lineage distribution in collaboration with other national and international laboratories.
1) Development of antivirals. In the frame of the EPIZONE project we have evaluated the effect of new biological antivirals based on siRNA. We have identified three regions on the nucleoprotein gene for the design of siRNA effective on the in vitro replication of PPRV but also RPV and MV. The inhibition of PPRV progeny is made by a factor of 104. In vivo therapeutic effect through viral and non-viral vectors are currently evaluated in a non-infectious animal model.
2) Development of a quantitative RT-PCR (QRT-PCR) for PPRV. In order to test the specificity and the sensitivity of the QRT-PCR for PPRV and other morbilliviruses, several strains including the vaccine strain, Nig PPR 75-1, were selected. Serial tenfold dilutions of the viral suspension were tested by classical RT-PCR and by QRT-PCR, to compare their sensitivity. The laboratory has also developed a robotized method (BIOMEK FX) for the viral RNA extraction. Samples from naturally infected small ruminants were used to validate the robotized extraction. This assay assessed the delectability of PPRV by real time RT-PCR in biological samples and confirmed the validity of the robotized extraction.
3) Improve knowledge of PPRV lineage distribution in the world. It is noted that expansion of PPR is linked to change in the current distribution of the genotypes of the virus. Molecular epidemiology has genetically divided PPR virus in four lineages; three historically settled in Africa and a single lineage, lineage IV, confined to Asia. Data collected over the last decade illustrate the wide incursion from the Middle East of lineage IV into Sudan and some neighbouring countries including the Central African Republic and Cameroun. The same lineage was found in Morocco in 2008. Two publications are related to this investigation (Banyard et al. 2010 and Kwiatek et al., accepted).
6.Collection, analysis and dissemination of epizootiological data relevant to international disease control
The laboratory is collecting, analysing and participating to the dissemination of epizootiological data through AU-IBAR and the Regional Laboratories Network organised for Africa. Countries are also sending individually samples from outbreaks for the diagnosis of PPR or RP.
Regarding OIE Member Countries, the notifications of PPR to OIE when positive results are found, is made by the country itself.
7.Provision of consultant expertise to OIE or to OIE Members
The Reference Laboratory expert provided advice directly to the OIE through an Ad hoc Group to review the OIE Pathway for rinderpest accreditation and to evaluate the country status.Attendance to working groups at the OIE or FAO Headquarters:
- Ad hoc group meeting for rinderpest accreditation and for evaluation of country status, OIE HQ, Paris, January 2010,
- Global Rinderpest Eradication Programme (GREP) Expert Consultation meeting, 12 – 15 October 2010,
- Second Global Conference of OIE Reference Laboratories and Collaborating Centres OIE Headquarters, Paris, France, 21–23 June 2010.
8.Provision of scientific and technical training to personnel from other OIE Members
Two PhD students from the following countries were hosted by CIRAD. The subjects of the different thesis are:
-Interfering RNA againstPeste des petits ruminants virus (PPRV) as a model for genetic-based therapy of morbilliviruses.
-In vitro dynamic of relapse mutants of peste des petits ruminants virus (PPRV) under RNA interference pressure: implication for anti-morbillivirus therapy.
- Epidemiology of peste des petits ruiminants in Sénégal.
Country/Diploma / Type of training / Starting date / lengthPakistan/PhD / Research / 01/01/10 / 11 months
Brasil/PhD / Research / 01/01/10 / 11 months
Côte d’Ivoire/Master 2 / Research / 01/03/10 / 10 months
9.Provision of diagnostic testing facilities to other OIE Members
Official diagnostic services were given to the following countries. Differentiation is made between tentative and confirmatory testing.
Country / Species / Sample / Number of tests / Number of positive samples/dubious / Test / Nature of the testConfirmatory or tentative
RPV2/PPRV1
Cattle / Serum / 129 / 0 / C-ELISA / Confirmatory
Brasil
Djibouti / Caprine/Camel / Serum / 16 / 8 / C-ELISA / Confirmatory
Senegal / Caprine/Ovine / Tissue / 24 / 16 / RT-PCR3/Sequence / Confirmatory
Caprine/Ovine / Tissue / 224 / 41 / RT-PCR3/Sequence / Confirmatory
Caprine/Ovine / Serum / 224 / 150/6 / C-ELISA / Confirmatory
Cameroon / Caprine / Serum / 34 / 2/4 / C-ELISA / Confirmatory
Djibouti / Dromedary / Serum / 2 / 0 / C-ELISA / Confirmatory
Oryx gazella beisa / Serum / 2 / 0 / C-ELISA / Confirmatory
Caprine / Serum / 10 / 8 / C-ELISA / Confirmatory
Mayetta / Caprine / Serum / 114 / 0 / C-ELISA / Confirmatory
Bovine / Serum / 1 / 0 / C-ELISA / Confirmatory
Caprine / Sang / 10 / 0 / RT-PCR/QPCR / Confirmatory
Bovine / Sang / 1 / 0 / RT-PCR/QPCR / Confirmatory
Caprine / lung / 2 / 0 / RT-PCR/QPCR / Confirmatory
Vaccine/Cell/Serum Contaminants
BVDV Cell/ Fœtal Calf Serum / 18 / QRT-PCR / Tentative
Jordany / - / PPR Vaccine / 2 / QualityControl4
Pass
1 PPRV: Peste des Petits Ruminants Virus.
2 RPV: Rinderpest Virus (all samples tested remained negative).
3 RT-PCR: Reverse Transcription and amplification.
4 Sterility test + PCR (RP, PPR, BVD, mycoplasma)+ titration (CPE visualized by immunoflorescence test using an anti-PPRV Mab)+ sequencing.
10.Organisation of international scientific meetings on behalf of OIE or other international bodies
None
11.Participation in international scientific collaborative studies
Collaboration with AU/IBAR, Nairobi, Kenya and the joint Division FAO/IAEA, Vienna, Austria on all aspects of the rinderpest network surveillance.
Participation to Epizone project (Network of Excellence for Epizootic Disease Diagnosis and Control)
which objective isto improve research on preparedness, prevention, detection, and control of epizootic diseases within Europe.
12.Publication and dissemination of information relevant to the work of OIE (including list of scientific publications, internet publishing activities, presentations at international conferences)
Presentations at international conferences and meetings
Libeau G. Potential implications of rinderpest eradication for peste des petits ruminants. FAO-GREP Symposium. Lessons learnt for the global rinderpest eradication 13 - 15 October 2010 Rome, Italy
Scientific publications in peer-reviewed journals
Ayari-Fakhfakh E, Ghram A, Bouattour A, Larbi I, Gribâa-Dridi L, Kwiatek O, Bouloy M, Libeau G, Albina E, Cêtre-Sossah C. First serological investigation of peste-des-petits-ruminants and Rift Valley fever in Tunisia. Vet J. 2010 Feb 16.
Banyard AC, Parida S, Batten C, Oura C, Kwiatek O, Libeau G. Global distribution of Peste des petits ruminants virus and prospects for improved diagnosis and control. J Gen Virol. 2010 Sep 15. [Epub ahead of print]
Keck N, Kwiatek O, Dhermain F, Dupraz F, Boulet H, Danes C, Laprie C, Perrin A, Godenir J, Micout L, Libeau G. Resurgence of Morbillivirus infection in Mediterranean dolphins off the French coast. Vet Rec. 2010 May 22;166(21):654-5. No abstract available. Erratum in: Vet Rec. 2010 Jun 19;166(25):1.
Khalafalla AI, Saeed IK, Ali YH, Abdurrahman MB, Kwiatek O, Libeau G, Obeida AA, Abbas Z. An outbreak of peste des petits ruminants (PPR) in camels in the Sudan. Acta Trop. 2010 Nov;116(2):161-5. Epub 2010 Aug 11.
Kwiatek O, Keita D, Gil P, Fernández-Pinero J, Jimenez Clavero MA, Albina E, Libeau G.Quantitative one-step real-time RT-PCR for the fast detection of the four genotypes of PPRV. J Virol Methods. 2010 May;165(2):168-77. Epub 2010 Jan 29.
Other communications
none
13.Inscription of diagnostic kits on the OIE Register
i)Did you participate in expert panels for the validation of candidate kits forinscription on the OIE Register? If yes, for which kits?
No
ii)Did you submit to the OIE candidate kits for inscriptionon the OIE Register? If yes, for which kits?
No
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 20101