OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory
Newcastle disease
OIE Reference Laboratory for Avian influenza
Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe),
Viale dell’Università,
10 – 35020 Legnaro (Padua),
ITALY
Tel.:
+39 049 8084371
Fax:
+39 049 8084360
e-mail address:
website:
icapua@izsvenezie.it
www.izsvenezie.it
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Dr Ilaria Capua
Director
OIE Reference Laboratory for Newcastle
Disease and Avian Influenza
Name(including Title and
Position) of OIE Reference
Expert:
Dr Ilaria Capua
Director
OIE Reference Laboratory for Newcastle
Disease and Avian Influenza
Name (including Title and
Position) of writer of this report
(if different from above):
Dr Calogero Terregino
Head of the Diagnostic Virology
Laboratory
OIE Reference Laboratory for Newcastle
disease and Avian influenza
Annual reports of OIE Reference Centres, 2011
1
Newcastle disease
Part I: Summary of general activities related to the disease
1.
Test(s) in use/or available for specified diseases at the Italian OIE ND laboratory
Table 1
2.
Test
For
Specificity
Total
Haemagglutination inhibition test (HI)
Antibody
Subtype
2863
Haemagglutination inhibition test (HI)
Viral typing
Subtype
18
Inoculation of SPF embryonated chicken eggs
Virus isolation
Group
856
rRT-PCR (reverse transcriptase- polymerase
chain reaction)
Virus
Group
3124
Sequencing
Virus
Pathotype
403
Production and distribution of diagnostic reagents
Table 2: Antigens produced in house at the Italian OIE ND laboratory in 2011
2
Antigen
ml produced
No of batches
H1N1
1720
16
H1N2
630
7
H2N1
100
1
H2N2
330
4
H2N3
100
1
H2N5
200
1
H2N6
100
1
H3N1
100
1
H3N2
570
7
H3N6
210
2
H3N8
2110
15
H5N1
4823
39
H5N2
343
3
H5N3
1150
4
H5N9
575
2
H7N1
1280
9
H7N3
910
5
H7N4
245
1
H7N7
2465
8
Annual reports of OIE Reference Centres, 2011
Newcastle disease
Antigen
ml produced
No of batches
H9N2
4566
49
H10N1
240
2
NDV
3242
12
IBV QX
17280
37
IBV M41
2350
6
IBV Q1
1380
5
IBV IS/1494/06
170
3
PR8
310
4
Negative
240
1
TOTAL
47,739
246
Table 3: Sera produced in house at the Italian OIE ND laboratory in 2011
Serum
ml produced
No of batches
Negative control serum
385
3
H1N1
123
2
H2N2
103
1
H2N1
103
1
H2N3
103
1
H3N2
164
1
H5N1
713
3
H5N2
385
4
H7N3
80
1
H9N2
604
6
H12N5
61
1
PMV1 (NDV)
332
3
PMV7
69
1
PMV8
217
1
IBV Q1
100
1
TOTAL
3,542
30
Table 4: Antigens supplied nationally and to laboratories in other countries by the Italian OIE ND laboratory in
2011
Antigen
Amount supplied (ml)
H1N1
106
H2N3
16
H3N2
4
H3N8
34
H4N8
16
Annual reports of OIE Reference Centres, 2011
3
Newcastle disease
4
Antigen
Amount supplied (ml)
H5N1
199
H5N2
358
H5N2 for AGID test
64
H5N3
182
H5N9
135
H6N2
45
H7N1
141
H7N3
262
H7N4
49
H7N7
171
H8N4
13
H9N2
80
H9N7
13
H10N1
22
H11N6
15
H11N9
3
H12N5
15
H13N6
16
H14N5
14
H15N9
14
H16N3
16
NDV
155
APMV1 (pigeon variant)
8
APMV2
10
APMV3
12
APMV4
9
APMV6
9
APMV7
13
APMV8
9
APMV9
9
NDV for ELISA test
16
IBV 624I
1
IBV QX
4
IBV D274
2
IBV 793B
2
IBV D1466
4
HEV
1
Annual reports of OIE Reference Centres, 2011
Newcastle disease
Antigen
Amount supplied (ml)
ILT for AGID test
5
TOTAL
2,272 ml (701 ml nationally and 1,571 ml to laboratories in
other countries)
Table 5: Sera supplied nationally and to laboratories in other countries by the Italian OIE ND laboratory in 2011
Sera
Amount supplied (ml)
Negative control serum
224
H1N1
45
H1N2
1
H2N3
19
H3N2
4
H3N8
22
H4N8
19
H5N1
93
H5N2
172
H5N3
100
H5N9
29
H6N2
37
H7N1
72
H7N3
49
H7N4
10
H7N7
120
H8N4
19
H9N2
57
H9N7
13
H10N1
19
H11N6
16
H11N9
8
H12N5
18
H13N6
20
H14N5
19
H15N9
17
H16N3
30
NDV
81
APMV1(pigeon variant)
7
APMV2
14
APMV3
15
Annual reports of OIE Reference Centres, 2011
5
Newcastle disease
Sera
Amount supplied (ml)
APMV4
11
APMV6
10
APMV7
11
APMV8
10
APMV9
10
IBV M41
2
IBV 624I
3
IBV D274
2
MG SAR
26
ILT for AGID test
3
TOTAL
1,457 ml (95 ml nationally and 1,362 ml to laboratories in
other countries)
Part II: 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
a)
Establishment and maintenance of a network with other OIE Reference Laboratories
designated for the same pathogen or disease and organisation of regular inter-laboratory
proficiency testing to ensure comparability of results
In March 2011, the Reference Laboratory participated in the European proficiency test on AI and ND
organized by the EU Reference Laboratory (AHVLA, Weybridge, UK) for EU member countries and some
extra-EU countries. The PT panel consisted of blind serum samples for the serological determination of the
avian influenza subtype/ND antibodies and antibody titre. In addition, blind samples for molecular testing
(RT-PCR and sequencing) were submitted. The laboratory achieved 100% of correct results.
b)
Organisation of inter-laboratory proficiency testing with laboratories other than OIE
Reference Laboratories for the same pathogens and diseases to ensure equivalence of
results
Ring trials organized by the Italian OIE Reference Laboratory during 2011
National Ring-test
In 2011 a serological and virological ring test was organised by the Italian National Reference Centre to
assess the capability of Italian public laboratories to identify selected antigens; to detect antibodies to avian
influenza viruses and Newcastle disease in avian sera; to standardize the diagnostic procedures used by the
different laboratories.
Each laboratory was identified with a number known to the laboratory itself only (stated upon consignment
of partial data).
As in the previous ring test, in 2011 each laboratory received 15 viral antigens (avian Orthomyxovirus and
Paramyxovirus) labelled with numbers from 1 to 15, and 15 sera for influenza virus and avian Paramyxovirus
labelled with letters from A to Q. Lyophilized reagents were also supplied.
All the laboratories were able to detect H5 and H7 subtypes viruses and NDV viruses by virological,
serological and molecular methods. Furthermore, most laboratories were able to detect the H1N1 pandemic
virus, although without specific reagents to detect this viral subtype, and an increased number of different
6
Annual reports of OIE Reference Centres, 2011
Newcastle disease
viral subtypes was identified.
The number of reagents will be increased in order to enable different laboratories to enlarge the diagnostic
efforts to identify influenza viruses in general.
International Ring-test
In addition to the National Ring-Test, the Reference Centre has organized the third Proficiency Test 2011
(PT 2011) for avian influenza and Newcastle disease for reference laboratories in different parts of the world
(North-Africa, West and Central Africa, South Africa, East Africa, Eastern Europe, Middle East and Central
Asia). This initiative was funded by UN-FAO (Food and Agriculture Organization of the United Nations).
A total of 48 countries were invitated to partecipate as follows:
All the laboratories were able to detect antibodies against H7 and H5 subtypes by HI test and to detect H5
viral RNA by molecular methods (RT-PCR and RRT-PCR). Areas of improvement have been identified in
the serological assay (AGID) and in molecular methods (e.g. RT-PCR and RRT-PCR) to identify H7
subtype viruses. Corrective actions are being implemented to improve the accuracy of these tests in specific
laboratories. This proficiency test was essential in providing the necessary feedback to FAO, donors and
reference laboratories to better design future training courses and provide technical support of the network.
23 laboratories of the Central Africa Regional Laboratory Network (RESOLAB), 8 laboratories of the
Eastern Africa Regional Laboratory Network (EARLN), two laboratories of the FAO-ECTAD Southern
Africa Regional Laboratory Network, four laboratories of the North Africa Laboratory Network, three
laboratories of the Central-Asia Laboratory Network, five laboratories of the Middle East area and three
laboratories of Eastern Europe countries. All the laboratories (100%) which sent results to the Reference
Centre perfomed the serological tests, while 18 laboratories (51%) performed the molecular tests. In detail,
the inibition of haemoagglutination (HI) was the most used test (97.1%), while AGID test was performed by
25 out of 35 laboratories (71.4%). Among the laboratories which perfomed the molecular tests, the most used
diagnostic protocol (14 out of 18 laboratories, 77.8%) was the diagnosis of H5 influenza subtype. Influenza
molecular diagnosis (Gene M) was performed by 13 out of 18 laboratories (72.2%), while the the diagnostic
protocol for detection of H7 subtype influenza virus was performed by 12 out of 18 (66.7%) laboratories. The
protocol for detection of ND was perfomed by 12 out of 18 laboratories (66,7%). The results of the PT 2011
have been presented during the closing meeting held in Bamako (Mali) in december 2011.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
See point 2 above.
5.
Research and development of new procedures for diagnosis and control
1)
Development of a Pyrosequencing-based method
Development of a Pyrosequencing-based method for the detection and differentiation of Avian
paramyxovirus type 1. Specificity, sensitivity and reproducibility of the method were assessed. The
sensitivity of the method ranged between 0.5x10 1.7 and 0.5x10 2.7 EID50/100 μl depending on the biological
specimens analyzed and the viral strain used. The results of the validation procedures indicate that the test is
suitable as a routine laboratory test for rapid detection and differentiation of class II APMV-1.
2)
Development of microneutralization test for Newcastle disease
The micro neutrialization technique for influenza has been modified for application to Newcastle disease.
The purpose is the evaluation in vitro of the response to vaccines compared to the tests currently in use
(ELISA, HI). The modifications are a different cell line (VERO cells instead of MDCK cells) and two
specific primary antibodies, that is monoclonal antibodies, directed to the surface glycoprotein of NDV. The
data obtained are currently under evaluation.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
None.
Annual reports of OIE Reference Centres, 2011
7
Newcastle disease
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
IZSVe operates in accordance with UNI CEI EN ISO/IEC 17025 and ISO 9001 guidelines. The procedures
referring to the diseases which fall within the mandate of the Reference Laboratory are accredited and the
qualifications are maintained by means of regular inspection visits by ACCREDIA – Italian Accreditation System
– the appointed Italian National Accreditation body. Accreditation guarantees that reports on testing, inspection
and certification of systems, products or personnel, are issued in compliance to the strictest international
requirements of conformity evaluation, including constant and rigorous surveillance of the behaviour of accredited
laboratories and bodies. The IZSVe’s facilities include a BL3 laboratory for manipulation of class 3 agents (ie:
HPAI H5N1 viruses) and other facilities for animal experiments. A new BL3 animal facility was completed in
2011 and a new BL3 laboratory will be completed by mid-2012.
8.
Provision of consultant expertise to OIE or to OIE Member Countries
During 2011, the Reference Laboratory Director provided advice in reviewing texts for the OIE, chapter 2.3.14 on
Newcastle disease of the Terrestrial Manual, responding to specific technical queries from OIE.
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
During 2011 the IZSVe continued and increased the training activities on avian influenza (AI) and Newcastle
disease (ND). In particular, it managed training courses on avian influenza diagnosis and epidemiology, for
veterinarians and technicians in partnership with international laboratories and other organizations – even private,
such as the Kyeema Foundation. The list of visitors is reported in Table 6.
Table 6: List of attendees of training courses at IZSVe during 2011
Name
Laboratory
Type of
Training
Activities
Kateri Bertran Dols
Centre de Recerca en Sanitat
Animal (CReSA), Spain
PhD
AI/ND classical and
molecular diagnostic
techniques
11/10/2010
31/01/2011
PhD
Diagnosis of AI/ND
and major avian
diseases
18/10/2010
30/04/2011
PhD
NDV Sequencing and
phylogenetic analysis,
epidemiology and
control
07/02/2011
18/02/2011
28/02/2011
11/03/2011
07/02/2011
10/07/2011
22/08/2011
26/08/2011
Department of Clinical Sciences
Hesamodin Emadi
Chashmi
Faculty of Veterinary Medicine,
Iran
Nagwa Osman Elshafie
College of Veterinary Medicine
and Animal Production,
Virology Department,
University of Science and
Technology, Khartoum, Sudan
Oumed Gergis
Mohamadamin
Ministry of Agriculture, Animal
Health Department, Kurdistan
Region, Erbil, Iraq
Private
ND and IBV: classical
and molecular
diagnosis;
epidemiology
Livia Victoria Patrono
Università degli Studi di
Bologna, Facoltà di Medicina
Veterinaria, Italy
Internship
AIV, NDV, IBV
Classical and
molecular diagnosis
and epidemiology
Lourenço Paulo Mapaco
Agricultural Research Institute
of Mozambique, Mozambique
Kyeema
Foundation
8
Laboratory biosafety
and quality assurance;
NDV classical and
molecular techniques
according to OIE/EU
Period
Annual reports of OIE Reference Centres, 2011
Newcastle disease
guidelines
Name
Laboratory
Raphael Samwel Sallu
Central Veterinary Laboratory,
Ministry of Livestock
Development and Fisheries,
Tanzania
Department of Animal Health
and Livestock Development,
Malawi
Julius L.C. Chulu
Musalwa Muyangwa
Ministry of Livestock and
Fisheries Development, Zambia
Type of
Training
Activities
Kyeema
Foundation
Laboratory biosafety
and quality assurance;
NDV classical and
molecular techniques
according to OIE/EU
guidelines
22/08/2011
26/08/2011
Kyeema
Foundation
Laboratory biosafety
and quality assurance;
NDV classical and
molecular techniques
according to OIE/EU
guidelines
22/08/2011
26/08/2011
Kyeema
Foundation
Laboratory biosafety
and quality assurance;
NDV classical and
molecular techniques
according to OIE/EU
guidelines
22/08/2011
26/08/2011
Period
10. Provision of diagnostic testing facilities to other OIE Member Countries
No. analyses
performed
(HI, ELISA,
AGID)
No. analyses
performed
(isolation in
embryonated
eggs/cell culture)
No. analyses
performed
(RRT/RT-PCR)
Primary diagnostic
(PD) Confirmatory
(C) testing
Total
AZERBAIJAN
36
4
20
16
PD
76
BANGLADESH
0
61
341
456
C
858
BULGARIA
0
0
2
0
C
2
BURKINA FASO
0
10
284
0
C
294
531
2
210
0
C
743
EGYPT
0
22
573
200
PD
795
JORDAN
0
6
71
24
PD
101
IRAN
0
15
68
567
C
650
ZIMBABWE
50
0
80
0
PD
130
Total
632
120
1727
752
Country
CAMEROON
No of analyses
performed
(sequencing)
3138
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
None.
12. Participation in international scientific collaborative studies
Annual reports of OIE Reference Centres, 2011
9
Newcastle disease
None.
13. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)
1.
Bonfante,F.; Heidari,A.; Salviato,A.; Monne,I.; Beato,M.S.; Raffini,E.; Taddei,R.; Biasini,G.; Terregino,C.
(2011); Caratterizzazione dei virus della malattia di Newcastle isolati da columbiformi selvatici in Italia. IV
Workshop Nazionale di Virologia Veterinaria; Brescia 09-10 Giugno 2011
2.
Cattoli,G.; Susta,L.; Terregino,C.; Brown,C. (2011); Newcastle disease: a review of field recognition and
current methods of laboratory detection. J Vet Diagn Invest; June 2011; epub
3.
Dundon,W.G.; Heidari,A.; Fusaro,A.; Monne,I.; Beato,M.S.; Cattoli,G.; Koch,G.; Starick,E.; Brown,I.H.;
Aldous,E.W.; Briand,F-X.; Le Gall-Reculé,G.; Jestin,V.; Jorgensen,P.; Berg,M.; Zohari,S.; Metreveli,G.;
Munir,M.; Stahl,K.; Albina,E.; Hammoumi,S.; Gil,P.; Servan de Almeida,R.; Smietanka,K.; DomaniskaBlicharz,K.; Minta,Z.; Van Borm,S.; Van Den Berg,T.; Martin,A.M.; Barbieri,I.; Capua,I. (2011); Genetic
data from avian influenza and avian paramyxoviruses generated by the European network of excellence
(EPIZONE) between 2006 and 2011 - Review and reccomendations for surveillance. Vet Microbiol 154;
209-221.
4.
Frasnelli,M.; Taddei,R.; Raffini,E.; Corazzari,V.; Fiorentini,L.; Tosi,G.; Fedrizzi,G.; Piro,R.; Lavazza,A.;
Gelmetti,D.; Bonfante,F.; Cotti,C.; Gelmini,L.; Terregino,C.; De Marco,M.A.; Delogu,M. (2011); Indagini su
un episodio di elevata mortalità in tortora dal collare orientale (streptopelia decaocto). IV Workshop
Nazionale di Virologia Veterinaria; Brescia 09-10 Giugno 2011
5.
Lupini,C.; Cecchinato,M.; Listorti,V.; Munoz,O.; Terregino,C.; Cecchettin,K.; Catelli,E. (2011); Interazione
fra vaccini vivi per la profilassi della rinotracheite del tacchino e della malattia di Newcastle somministrati in
associazione ad un giorno di vita nel tacchino; Proceeding of 50th Meeting Forlì 7-8 Aprile 2011
_______________
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Annual reports of OIE Reference Centres, 2011