Abstract
By removing the N-terminal hydrophobic sequence, truncated VP2 (tVP2) genes were cloned into the pET-32a (+) plasmid and subsequently expressed as His fusion proteins. The puriï¬ed recombinant tVP2 proteins were speciï¬c to canine parvovirus (CPV), and one of them was used in an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of CPV antibodies. The minimum detection limit of this method was 1:1280. There was good agreement between tVP2-based indirect ELISA and the commercially available diagnostic kit. The results suggest that the recombinant tVP2 protein-based ELISA could be used to detect CPV antibodies. Key words: Canine parvovirus, recombinant truncated VP2 (tVP2), enzyme-linked immunosorbent assay (ELISA), antibody detection
Highlights
Canine parvovirus (CPV) is classified within the parvoviridae family
The results suggest that the recombinant truncated VP2 (tVP2) protein-based enzymelinked immunosorbent assay (ELISA) could be used to detect canine parvovirus (CPV) antibodies
To obtain the fusion protein, two truncated VP2 genes were expressed by using E. coli BL21 and plasmid vector pET-32a (+)
Summary
Canine parvovirus (CPV) is classified within the parvoviridae family. The CPV genome is a linear single-stranded DNA of approximately 5 kb, making it one of the smallest animal DNA viruses (Chapman and Rossmann, 1993). Dogs infected with CPV may develop acute bloody diarrhea, fever, and dehydration, and these symptoms are sometimes followed by shock and sudden death (Carman and Povey, 1985). CPV was recognized in 1978 and is considered endemic in virtually all populations of domestic and wild canines. It causes substantial emotional hardships to the animal owner, and often causes significant suffering in infected dogs. It would be beneficial to develop a fast and available diagnostic method to evaluate the maternally derived antibodies and the active response to pup vaccinetion (Decaro and Buonavoglia, 2012)
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