- S. Blome (4)
- P. Bonulauri (1)
- Thomas Bruun Rasmussen (2)
- Anette Bøtner (2)
- Akbar Dastjerdi (2)
- P.L. Eble (1)
- J. Fernández-Pinero (1)
- Y. Geurts (1)
- Béatrice Grasland (2)
- I. Greiser-Wilke (1)
- Wim H.M. Poel Van Der (1)
- A. Haegeman (2)
- Dennis Hanke (2)
- B. Hoffmann (1)
- M. Hofmann (1)
- M.A. Hofmann (1)
- M.M. Hulst (1)
- Marcel Hulst (1)
- M. Isaksson (1)
- Graham J. Belsham (2)
- F. Koenen(older publications) (1)
- F. Koenen (3)
- E. Lange (1)
- Antonio Lavazza (2)
- N. Leblanc (1)
- I. Leifer (2)
- W.L.A. Loeffen (3)
- W. Loeffen (1)
- H.W.M. Moonen-Leusen (1)
- Alice Papetti (2)
- Jean Pierre Frossard (2)
- W.H.M. Poel van der (2)
- Anne Pohlmann (2)
- M.F. Potier Le (1)
- J. Quak (1)
- T.B. Rasmussen (1)
- T. Rosen von (1)
- S. Schroeder (1)
- T. Stadejek (1)
- K. Stahl (1)
- Falko Steinbach (2)
- M. Tignon (1)
- A. Uttenthal (2)
Sequence diversity of CV777 PEDV strains
Rasmussen, Thomas Bruun ; Boniotti, Maria Beatrice ; Papetti, Alice ; Grasland, Béatrice ; Frossard, Jean Pierre ; Dastjerdi, Akbar ; Hulst, M.M. ; Hanke, Dennis ; Pohlmann, Anne ; Blome, Sandra ; Poel, W.H.M. van der; Steinbach, Falko ; Blanchard, Yannick ; Lavazza, Antonio ; Bøtner, Anette ; Belsham, Graham J. - \ 2018
PRJEB20818 - ERP023004
Full-length genome sequences of porcine epidemic diarrhoea virus strain CV777; use of NGS to analyse genomic and sub-genomic RNAs
Rasmussen, Thomas Bruun ; Boniotti, Maria Beatrice ; Papetti, Alice ; Grasland, Béatrice ; Frossard, Jean Pierre ; Dastjerdi, Akbar ; Hulst, Marcel ; Hanke, Dennis ; Pohlmann, Anne ; Blome, Sandra ; Poel, Wim H.M. Van Der; Steinbach, Falko ; Blanchard, Yannick ; Lavazza, Antonio ; Bøtner, Anette ; Belsham, Graham J. - \ 2018
PLoS One 13 (2018)3. - ISSN 1932-6203
Porcine epidemic diarrhoea virus, strain CV777, was initially characterized in 1978 as the causative agent of a disease first identified in the UK in 1971. This coronavirus has been widely distributed among laboratories and has been passaged both within pigs and in cell culture. To determine the variability between different stocks of the PEDV strain CV777, sequencing of the full-length genome (ca. 28kb) has been performed in 6 different laboratories, using different protocols. Not surprisingly, each of the different full genome sequences were distinct from each other and from the reference sequence (Accession number AF353511) but they are >99% identical. Unique and shared differences between sequences were identified. The coding region for the surface-exposed spike protein showed the highest proportion of variability including both point mutations and small deletions. The predicted expression of the ORF3 gene product was more dramatically affected in three different variants of this virus through either loss of the initiation codon or gain of a premature termination codon. The genome of one isolate had a substantially rearranged 5´-terminal sequence. This rearrangement was validated through the analysis of sub-genomic mRNAs from infected cells. It is clearly important to know the features of the specific sample of CV777 being used for experimental studies.
Efficacy of chimeric Pestivirus vaccine candidates against Classical Swine Fever: protection and DIVA characteristics
Eble, P.L. ; Geurts, Y. ; Quak, J. ; Moonen-Leusen, H.W.M. ; Blome, S. ; Hofmann, M.A. ; Koenen, F. ; Beer, M. ; Loeffen, W.L.A. - \ 2013
Veterinary Microbiology 162 (2013)2-4. - ISSN 0378-1135 - p. 437 - 446.
subunit marker vaccine - hog-cholera virus - monoclonal-antibodies - pigs - transmission - differentiation - strains - infection - virulent - induce
Currently no live DIVA (Differentiating Infected from Vaccinated Animals) vaccines against classical swine fever (CSF) are available. The aim of this study was to investigate whether chimeric pestivirus vaccine candidates (CP7_E2alf, Flc11 and Flc9) are able to protect pigs against clinical signs, and to reduce virus shedding and virus transmission, after a challenge with CSF virus (CSFV), 7 or 14 days after a single intramuscular vaccination. In these vaccine candidates, either the E2 or the Erns encoding genome region of a bovine viral diarrhoea virus strain were combined with a cDNA copy of CSFV or vice versa. Furthermore, currently available serological DIVA tests were evaluated. The vaccine candidates were compared to the C-strain. All vaccine candidates protected against clinical signs. No transmission to contact pigs was detected in the groups vaccinated with C-strain, CP7_E2alf and Flc11. Limited transmission occurred in the groups vaccinated with Flc9. All vaccine candidates would be suitable to stop on-going transmission of CSFV. For Flc11, no reliable differentiation was possible with the current Erns-based DIVA test. For CP7_E2alf, the distribution of the inhibition percentages was such that up to 5% false positive results may be obtained in a large vaccinated population. For Flc9 vaccinated pigs, the E2 ELISA performed very well, with an expected 0.04% false positive results in a large vaccinated population. Both CP7_E2alf and Flc9 are promising candidates to be used as live attenuated marker vaccines against CSF, with protection the best feature of CP7_E2alf, and the DIVA principle the best feature of Flc9.
|Evaluation of classical swine fever virus antibody detection assays with an emphasis on the differentiation of infected from vaccinated animals
Schroeder, S. ; Rosen, T. von; Blome, S. ; Loeffen, W.L.A. ; Haegeman, A. ; Koenen, F. ; Uttenthal, A. - \ 2012
Revue scientifique et technique / Office International des Epizooties 31 (2012)3. - ISSN 0253-1933 - p. 997 - 1010.
The aim of this study was to evaluate the general characteristics of
commercially available enzyme-linked immunosorbent assays (ELISAs) to detect
antibody against classical swine fever (CSF), as well as to assess their potential
use as accompanying marker tests able to differentiate infected from vaccinated
The Chekit* CSF-Sero and the HerdChek* CSFV Ab, both of which detect
antibodies against the E2 protein of classical swine fever virus (CSFV), had the
highest sensitivity. Both tests were practicable and showed good reproducibility.
Comparable sensitivity was shown by the Chekit* CSF-Marker, an Erns ELISA.
However, this test does not allow differentiation between antibodies directed
against ruminant pestiviruses and those against CSFV. Therefore, it is not
suitable for use with the chimeric marker vaccines tested.
The PrioCHECK® CSFV Erns was the only ELISA suitable for use in DIVA with
marker vaccines containing Erns proteins from ruminant pestiviruses. However,
this test was less sensitive and selective than the E2-ELISAs and cannot be
Comparative evaluation of live marker vaccine candidates "CP7_E2alf" and "flc11" along with C-strain "Riems" after oral vaccination
Blome, S. ; Aebischer, A. ; Lange, E. ; Hofmann, M. ; Leifer, I. ; Loeffen, W.L.A. ; Koenen, F. ; Beer, M. - \ 2012
Veterinary Microbiology 158 (2012)1-2. - ISSN 0378-1135 - p. 42 - 59.
classical-swine-fever - time rt-pcr - e-rns - virus - pestiviruses - differentiation - pigs - e2
Due to the tremendous socio-economic impact of classical swine fever (CSF) outbreaks, emergency vaccination scenarios are continuously under discussion. Unfortunately, all currently available vaccines show restrictions either in terms of marker capacities or immunogenicity. Recent research efforts were therefore directed at the design of new modified live marker vaccines. Among the most promising candidates the chimeric pestiviruses “CP7_E2alf” and “flc11” were identified. Within an international research project, these candidates were comparatively tested in challenge experiments after a single oral vaccination. Challenge infection was carried out with highly virulent CSF virus strain “Koslov”, 14 or 21 days post vaccination (dpv), respectively. Safety, efficacy, and marker potential were addressed. All assessments were done in comparison with the conventional “gold standard” C-strain “Riems” vaccine. In addition to the challenge trials, multiple vaccinations with both candidates were performed to further assess their marker vaccine potential. All vaccines were safe and yielded full protection upon challenge 21 days post vaccination. Neither serological nor virological investigations showed major differences among the three vaccines. Whereas CP7_E2alf also provided clinical protection upon challenge at 14 days post vaccination, only 50% of animals vaccinated with flc11, and 83% vaccinated with C-strain “Riems” survived challenge at this time point. No marked differences were seen in protected animals. Despite the fact that all multiple-vaccinated animals stayed sero-negative in the accompanying marker test, the discriminatory assay remains a weak point due to delayed or inexistent detection of some of the vaccinated and subsequently infected animals. Nevertheless, the potential as live marker vaccines could be confirmed for both vaccine candidates. Future efforts will therefore be directed at the licensing of “Cp7_E2alf” as the first live marker vaccine for CSF
Classical swine fever virus detection: results of a real-time reverse transcription polymerase chain reaction ring trial conducted in the framework of the European network of excellence for epizootic disease diagnosis and control.
Hoffmann, B. ; Blome, S. ; Bonulauri, P. ; Fernández-Pinero, J. ; Greiser-Wilke, I. ; Haegeman, A. ; Isaksson, M. ; Koenen, F. ; Leblanc, N. ; Leifer, I. ; Potier, M.F. Le; Loeffen, W. ; Rasmussen, T.B. ; Stadejek, T. ; Stahl, K. ; Tignon, M. ; Uttenthal, A. ; Poel, W.H.M. van der - \ 2011
Journal of Veterinary Diagnostic Investigation 23 (2011)5. - ISSN 1040-6387 - p. 999 - 1004.
rt-pcr assay - pestiviruses - validation - infection - virulence - vaccine - taqman - probes - signs - pigs
The current study reports on a real-time reverse transcription polymerase chain reaction (real-time RT-PCR) ring trial for the detection of Classical swine fever virus (CSFV) genomic RNA undertaken by 10 European laboratories. All laboratories were asked to use their routine in-house real-time RT-PCR protocols and a standardized protocol commonly used by the Friedrich-Loeffler-Institute (FLI) on a panel of well-characterized samples. In general, all participants produced results within the acceptable range. The FLI assay, several in-house assays, and the commercial kits had high analytical sensitivity and specificity values. Nevertheless, some in-house systems had unspecific reactions or suboptimal sensitivity with only a single CSFV genotype. Follow-up actions involved either improvement of suboptimal assays or replacement of specific laboratory assays with the FLI protocol, with or without modifications. In conclusion, the ring trial showed reliability of classical swine fever diagnosis on an international level and helped to optimize CSFV-specific RT-PCR diagnostics.