An ELISA based on the repeated foot-and-mouth disease virus 3B epitope peptide can distinguish infected and vaccinated cattle |
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Authors: | Mingchun?Gao Runxiang?Zhang Meng?Li Shuang?Li Yongsheng?Cao Bo?Ma Email author" target="_blank">Junwei?WangEmail author |
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Institution: | (1) College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People’s Republic of China; |
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Abstract: | To develop a strategy of differentiating infected from vaccinated animals (DIVA) with foot-and-mouth disease virus (FMDV),
a short (27aa) peptide containing three conserved linear B cell epitopes of the FMDV 3B nonstructural protein was designed.
This novel BF peptide was synthesized using a gene splicing by overlap extension protocol with preferred codons for Escherichia coli. The resultant eight tandem repeat multimer (1, 2, 4, 6, 8, 16, 24, and 32BF) were expressed as soluble fusion proteins in
E. coli. An indirect ELISA was developed based on the recombinant 8BF protein with the aim of specifically distinguishing antibodies
induced by FMDV infection but not those induced by vaccination. Using the cut-off value of 0.3, the sensitivity of the assay
was 96.8% and the specificities for naive and vaccinated cattle were 99.8 and 99.0%, respectively. The performance of the
newly developed epitope-based ELISA was compared with three commercial NSP ELISA kits. The 8BF-ELISA appears to be a promising
DIVA test for FMD control and eradication. |
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