OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Rinderpest and Peste des petits ruminants
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:
genevieve.libeau@cirad.fr
http://www.cirad.
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
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
1
Rinderpest and Peste des petits ruminants
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
Total
ELISA
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-specific
PPR-specific
0
2
1
100
1
100
RT-PCR
Genome
RP-specific
PPR-specific
Pan-morbillivirus
DVM
261
261
261
0
100
100
0
0
361
361
261
0
QRT-PCR
Genome
RP-specific
PPR-specific
Pan-morbillivirus
0
100
0
0
600
0
0
700
0
Peptide ELISA
Antibody
PPR-specific
0
0
0
Sequencing
Genome
RP
PPR
DMV
0
30
0
1
10
0
1
40
Cell culture
Virus isolation
RP
PPR
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 CELISA 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 (http://www.oie.int/eng/normes/mcode/en_chapitre_1.1.6.htm) 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...
2
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Rinderpest and Peste des petits ruminants
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, interlaboratory ring trials for PPR antibody detection were organized. 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
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
3
Rinderpest and Peste des petits ruminants
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:
8.

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.
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:
4
-
Interfering RNA against Peste 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
length
Pakistan/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
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Rinderpest and Peste des petits ruminants
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 test
Confirmatory or
tentative
RPV2/PPRV1
Brasil
Cattle
Serum
129
0
C-ELISA
Confirmatory
Djibouti
Caprine/Camel
Serum
16
8
C-ELISA
Confirmatory
Senegal
Caprine/Ovine
Tissue
24
16
RTPCR3/Sequence
Confirmatory
Caprine/Ovine
Tissue
224
41
RTPCR3/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
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
Mayetta
Vaccine/Cell/Serum Contaminants
BVDV Cell/ Fœtal Calf Serum
Jordany
-
QRTPCR
18
PPR Vaccine
2
1
PPRV : Peste des Petits Ruminants Virus.
2
RPV : Rinderpest Virus (all samples tested remained negative).
QualityControl4
Tentative
Pass
3
RT-PCR : Reverse Transcription and amplification.
Sterility test + PCR (RP, PPR, BVD, mycoplasma)+ titration (CPE visualized by immunoflorescence test using
an anti-PPRV Mab)+ sequencing.
4
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.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
5
Rinderpest and Peste des petits ruminants
Participation to Epizone project (Network of Excellence for Epizootic Disease Diagnosis and Control)
which objective is to 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):16877. 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 for inscription on the
OIE Register? If yes, for which kits?
No
ii)
Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for
which kits?
No
_______________
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010