Journal
VETERINARY MICROBIOLOGY
Volume 161, Issue 3-4, Pages 274-285Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.vetmic.2012.07.046
Keywords
Classical Swine Fever virus; Pestivirus; DIVA; CP7_E2alf; Discriminatory ELISA; Marker vaccine
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Funding
- European Community [227003 CP-FP (CSFV_goDIVA)]
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New generations of Classical Swine Fever virus (CSFV) marker vaccines have recently been developed in order to make emergency vaccination in case of a CSF outbreak more feasible. However, the application of a marker vaccine is dependent on the availability of an accompanying discriminatory test allowing differentiation of infected from vaccinated animals (DIVA). CP7_E2alf, the most promising live marker vaccine candidate currently available, is a genetically modified Bovine Viral Diarrhea virus expressing the E2 glycoprotein of CSFV strain Alfort/187. The DIVA principle going along with CP7_E2alf is based on the detection of CSFV E-rns-specific antibodies that are raised in the host upon CSFV infection but not after vaccination with the marker vaccine. The aim of this study was to develop novel DIVA tests to be used in combination with CP7_E2alf. Two indirect ELISAs (one for screening, the other one for confirmation purposes) using bacterially expressed recombinant Ems proteins were designed and evaluated. Both ELISAs detected CSFV-specific antibodies against a broad range of strains and genotypes, and as early as 10 days after infection. They were able to distinguish CSFV-infected pigs from pigs vaccinated with CP7_E2alf and allowed discrimination of antibodies against ruminant pestiviruses in both, sera from domestic pigs and wild boar. Sensitivity and specificity of the screening ELISA was >= 95%. Thus, the ELISAs represent promising DIVA diagnostic tools, as well as an alternative to traditional pestivirus antibody differentiation by serum neutralization test. (C) 2012 Elsevier B.V. All rights reserved.
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