Development of a Competitive Enzyme-Linked Immunosorbent Assay Based on Purified Recombinant Viral Protein 7 for Serological Diagnosis of Epizootic Haemorrhagic Disease in Camels
Author(s) -
Anna Serroni,
Simonetta Ulisse,
Mariangela Iorio,
Caterina Laguardia,
Lilia Testa,
Gisella Armillotta,
Marco Caporale,
Romolo Salini,
Davide Lelli,
U. Wernery,
R. Raghavan,
Maria Teresa Mercante,
Mauro Di Ventura
Publication year - 2022
Publication title -
journal of tropical medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.747
H-Index - 30
eISSN - 1687-9694
pISSN - 1687-9686
DOI - 10.1155/2022/5210771
Subject(s) - virology , serology , biology , recombinant dna , orbivirus , epizootic , serotype , immunoassay , antibody , reoviridae , virus , seroprevalence , immunology , rotavirus , biochemistry , gene
Epizootic haemorrhagic disease virus (EHDV) is a member of the Orbivirus genus in the Reoviridae family, and it is the etiological agent of an arthropod-transmitted disease that affects domestic and wild ruminants. Due to its significant economic impact, many attempts have been done in order to develop diagnostic immunoassays mainly based on the use of the viral protein 7 (VP7), that is, the immunodominant serogroup-specific antigen. In this work, a recombinant VP7 (recVP7) of EHDV serotype 2 was produced in a baculovirus system, and after purification using ion metal affinity chromatography, we obtained a high yield of recombinant protein characterized by a high degree of purity. We used the purified recVP7 as reagent to develop a competitive enzyme-linked immunoassay (c-ELISA), and we tested the presence of EHDV antibodies in 185 dromedary camel serum samples. The c-ELISA showed good performance parameters in recognising positive sera of naturally EHDV-infected dromedary camels; in particular, our developed test reached 85.7% of sensitivity, 98.1% of specificity, 93% of accuracy, and a high agreement value with results obtained by the commercial ELISA kit (Cohen’s kappa value of 0.85) that we adopted as the reference method. This c-ELISA could be a useful screening test to monitor the virus spread in camels that are sentinel animals for endemic areas of disease.
Accelerating Research
Robert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom
Address
John Eccles HouseRobert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom