Abstract
BackgroundIn Poland, the leader in goose production in Europe, goose parovirus infection, or Derzsy’s disease (DD), must be reported to the veterinary administration due to the serious economic and epizootic threat to waterfowl production. Prophylactic treatment for DD includes attenuated live or inactivated vaccines. Moreover, the control of DD includes the monitoring of maternal derived antibody (MDA) levels in the offspring and antibody titers in the parent flock after vaccination. The aim of this study was to develop an ELISA for the detection of goose parvovirus (GPV) antibodies.ResultsTwo recombinant protein fragments derived from VP3 (viral protein 3) GPV, namely VP3ep6 and VP3ep4–6 with a mass of 20.9 and 32.3 kDa, respectively, were produced using an Escherichia coli expression system. These proteins were purified by one-step nickel-affinity chromatography, which yielded protein preparations with a purity above 95%. These recombinant proteins were useful in the detection of serum anti-GPV antibodies, and this was confirmed by Western blotting. However, recombinant VP3ep4–6 protein showed a greater ability to correctly identify sera from infected geese. In the next stage of the project, a pool of 166 goose sera samples, previously examined by a virus neutralization test (VN), was tested. For further studies, one recombinant protein (VP3ep4–6) was selected for optimization of the test conditions. After optimization, the newly developed ELISA was compared to other serological tests, and demonstrated high sensitivity and specificity.ConclusionIn conclusion, the VP3ep4–6 ELISA method described here can be used for the detection of antibodies to GPV in serum.
Highlights
In Poland, the leader in goose production in Europe, goose parovirus infection, or Derzsy’s disease (DD), must be reported to the veterinary administration due to the serious economic and epizootic threat to waterfowl production
Expression and purification of the goose parvovirus (GPV) VP3 recombinant proteins Two GPV VP3 recombinant proteins (VP3ep4–6 amino acid 311–534 and VP3ep6 amino acid 418–534) containing six histidyl residues at N- and C-terminals were expressed as soluble proteins with a calculated molecular mass of 32.3 kDa and 20.9 kDa, respectively (Fig. 1)
Purification of the GPV recombinant proteins was accomplished with a one-step chromatography procedure using metal-affinity chromatography with Ni2+ bound to iminodiacetic acidagarose (Novagen) and resulted in a production yield of about 40 mg and 30 mg of purified proteins (VP3ep4–6 and VP3ep6, respectively) per liter of cell culture
Summary
In Poland, the leader in goose production in Europe, goose parovirus infection, or Derzsy’s disease (DD), must be reported to the veterinary administration due to the serious economic and epizootic threat to waterfowl production. The aim of this study was to develop an ELISA for the detection of goose parvovirus (GPV) antibodies. Goose parvovirus (GPV) infection, known as Derzsy’s disease (DD), is a contagious, acute gastrointestinal disease that occurs in geese (Anser anser) and Muscovy ducks (Cairina moschata). The virus has a high mortality rate of up to 100% and causes a significant decrease in weight, loss of feathers on the back and neck and diarrhea [1]. It is severe for young goslings and Muscovy ducklings with maternal antibody deficiency. Despite widespread vaccination against GPV in the last 30 years, epidemiological data indicates its constant presence [12]
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