OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Infectious bovine rhinotracheitis/
Infectious pustular vulvovagenitis
Institute of Diagnostic Virology
FRIEDRICH-LOEFFLER-INSTITUT
Südufer 10
D-17493 Greifswald-Insel Riems
GERMANY
Tel:
(+49-383) 517.1223
Fax:
(+49-383) 517.1275
e-mail address:
website:
Martin.Beer@fli.bund.de
www.fli.bund.de
Name of Head of Laboratory
(Responsible Official):
PD Dr Martin Beer
Name of OIE Reference Expert:
PD Dr Martin Beer
Name of writer of this report
(if different from above):
Annual reports of OIE Reference Laboratories and Collaborating Centres, 20010
1
Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis
Summary of general activities related to the disease
1.
Test(s) in use/or available for the specified disease at your laboratory
Test
For
Specificity
Total
Indirect ELISA
BoHV-1-antibody
(serum, plasma, milk)
BoHV-1
464
gB-blocking-ELISA
BoHV-1-gB-antibody
(serum, plasma)
BoHV-1
1169
gE-blocking-ELISA
BoHV-1-gE-antibody, marker test
(serum, plasma)
wild type BoHV-1
2534
Neutralisation test (NT)
Plaque reduction test
BoHV-1-antibody (serum)
BoHV-1
HV
29
94
Cell culture
(MDBK, KOP-R)
Virus isolation (semen, swabs, organ
samples, vaccines)
Replication competent
virus
9
IFAT
BoHV-1 antigen (cell culture)
BoHV-1
9
PCR
Genome of BoHV-1 (semen, swabs,
organ samples, vaccines)
BoHV-1
63
Genome of herpesviruses
HV
46
Genome differentiation of bovine
herpesviruses (virus isolates)
HV
1
REFLP
2.
Production and distribution of diagnostic reagents
10 reference sera were collected from calves experimentally infected with BHV-2 as reference for studying cross
reactivity with BoHV-1. Approximately 20 milk samples (0.5-2 l) were obtained from BoHV-1 field-infected
cattle. In total, more than 150 reference samples (serum and milk) are now available.
103 reference sera and 45 reference milk samples were supplied nationally to the regional laboratories, and
225 reference samples (serum: 140; milk: 85) were supplied to other OIE member countries.
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
60 commercial BoHV-1-ELISAs were tested for batch release by the OIE reference laboratory. One new BHV-1
gB-blocking ELISA test was examined for licensing on the German market.
A novel triplex real-time PCR assay for the rapid detection and differentiation of field and vaccine strains of
bovine herpesvirus type 1 was validated and published. A broad series of swab samples, semen and tissues were
examined as well as a panel of ruminant herpesviruses.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis
41 pairs of trigeminal ganglia from cattle and additional diagnostic samples were tested for the presence of
genome sequences of BoHV-1 or of other ruminant herpesviruses by a BoHV-1 specific, newly designed real-time
PCR (gD, gE), and a pan-herpes PCR which detects conserved sequences of the herpesviral DNA-dependent
DNA-polymerase. The tests are currently applied to elucidate potential introduction of ruminant
alphaherpesviruses into cattle herds.
The OIE reference laboratory participated in a BoHV-1 proficiency test organised by the COORDINATION
CENTRE FOR VETERINARY DIAGNOSTICS (CCVD, Brussels, Belgium). 100% of the serum samples were
correctly characterised for the presence of BoHV-1 gE- and gB-specific antibodies.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
The German BoHV-1 reference sera R1 (positive), R2 (weak positive), R3 (very weak positive), R31 (negative)
and R32 (negative) were supplied to laboratories in Germany, France, Switzerland, Turkey and Morroco.
5.
Research and development of new procedures for diagnosis and control
Introduction of various ruminant alphaherpesviruses into cattle herds is under discussion to evolve as a threat to
advanced BoHV-1 eradication programs. High graded serological cross reactivity with BoHV-1 is observed in
numerous ELISA tests and even in serum neutralisation assays. Therefore, BoHV-1 antibody negative calves were
experimentally inoculated with BoHV-2 to elucidate whether a cross reactivity in BoHV-1 ELISA tests could be
induced. Serological cross reactivity between BoHV-2 and BoHV-1 might explain epidemiologically some of the
non plausible reactions in BoHV-1-free areas in Southern Germany where BoHV-2 is highly prevalent.
South American camelids were intranasally infected with BoHV-1 to study a potential role of llamas and alpacas
in BoHV-1 epidemiology.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
Data concerning the progress of the German BoHV-1 eradication program were collected and analysed using a
detailed questionnaire. In summary, more than 87% of the cattle holdings in Germany were BoHV-1-antibody or
BoHV-1-gE-antibody free at the end of 2009.
7.
Provision of consultant expertise to OIE or to OIE Members
Protocols, e.g. for BoHV-1-neutralization assays and BoHV-1 PCRs, were transferred to several international
laboratories. Expert advices concerning BoHV-1 diagnostics and BoHV-1 eradication have been required by
colleagues from UK, Spain, India, and Italy.
8.
Provision of scientific and technical training to personnel from other OIE Members
Not applicable
9.
Provision of diagnostic testing facilities to other OIE Members
Buffalo trigeminal samples from Italy were transferred to the FLI for differentiation of BoHV-1 and other
ruminant herpesviruses (BuHV-1), and bovine trigeminal ganglia were screened for the presence of latent BoHV-1
after experimental infection of cattle with BuHV-1. Buffalo serum samples from Switzerland were tested for the
presence of BoHV-1 specific antibodies.
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
Not applicable
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis
11. Participation in international scientific collaborative studies
Not applicable
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
Kerstin Wernike, Bernd Hoffmann, Donata Kalthoff, Patricia König, Martin Beer. Development and validation of
a triplex real-time PCR assay for the rapid detection and differentiation of field and vaccine strains of Bovine
Herpesvirus type 1. 4th Annual Meeting of EPIZONE, Saint Malo, France, 2010.
Patricia König. Perspectives and limitations of the pan herpes Dpol PCR. Meeting of the German association of
veterinary diagnosticians (AVID) in Kloster Banz, Germany, 2010.

Scientific publications in peer-reviewed journals
Kalthoff D, König P, Trapp S, Beer M. 2010. Immunization and challenge experiments with a new modified live
bovine herpesvirus type 1 marker vaccine prototype adjuvanted with a co-polymer. Vaccine, 28(36):5871-7.

Other communications
M. Beer revised and updated the chapter on IBR/IPV in the OIE Manual.
Beer, M, Koenig, P, and Höreth-Böntgen, D. 2009. BHV-1. Tiergesundheitsjahrbuch 2009, Friedrich-LoefflerInstitut, Germany.
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
_______________
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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