title: Development of a Luminex-Based DIVA Assay for Serological Detection of African Horse Sickness Virus in Horses.
abstract: African horse sickness (AHS) is considered a fatal re-emergent vector-borne disease of horses. In the absence of any effective treatment for AHS, vaccination remains the most effective form of disease control. The new generation of vaccines, such as one based on purified, inactivated AHS virus (AHSV, serotype 4), which does not induce antibodies against non-structural protein 3 (NS3), enables the development of diagnostic methods that differentiate infected from vaccinated animals (DIVA assays). As detecting AHS in AHSV-free countries may lead to restrictions on international animal movements and thereby cause significant economic damage, these DIVA assays are crucial for reducing movement restrictions. In this article, we describe a Luminex-based multiplex assay for DIVA diagnosis of AHS, and we validate it in a duplex format to detect antibodies against structural protein 7 (VP7) and NS3 in serum samples from horses vaccinated with inactivated AHSV4 vaccine or infected with a live virus of the same serotype. Results of the Luminex-based assay for detecting anti-NS3 antibodies showed good positive correlation with results from an in-house enzyme-linked immunosorbent assay (ELISA). Thus, the Luminex-based technique described here may allow multiplex DIVA antibody detection in a single sample in less than 2 h, and it may prove adaptable for the development of robust, multiplex serological assays.
authors.affiliation: VISAVET Health Surveillance Centre, Universidad Complutense Madrid, Madrid, Spain.|Animal Health Department, Universidad Complutense Madrid, Madrid, Spain.|CEI Campus Moncloa, UCM-UPM, Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Sánchez-Matamoros
authors.affiliation: VISAVET Health Surveillance Centre, Universidad Complutense Madrid, Madrid, Spain.|Animal Health Department, Universidad Complutense Madrid, Madrid, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Nieto-Pelegrín
authors.affiliation: Animal Health Laboratory, French Agency for Food, Environmental and Occupational Health and Safety (ANSES), UPE, UMR 1161 ANSES, INRA, ENVA, Maisons-Alfort, France.
authors.forename: C
authors.initials: C
authors.lastname: Beck
authors.affiliation: VISAVET Health Surveillance Centre, Universidad Complutense Madrid, Madrid, Spain.|Animal Health Department, Universidad Complutense Madrid, Madrid, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Rivera-Arroyo
authors.affiliation: Animal Health Laboratory, French Agency for Food, Environmental and Occupational Health and Safety (ANSES), UPE, UMR 1161 ANSES, INRA, ENVA, Maisons-Alfort, France.
authors.forename: S
authors.initials: S
authors.lastname: Lecollinet
authors.affiliation: Animal Health Laboratory, French Agency for Food, Environmental and Occupational Health and Safety (ANSES), UPE, UMR 1161 ANSES, INRA, ENVA, Maisons-Alfort, France.
authors.forename: C
authors.initials: C
authors.lastname: Sailleau
authors.affiliation: Animal Health Laboratory, French Agency for Food, Environmental and Occupational Health and Safety (ANSES), UPE, UMR 1161 ANSES, INRA, ENVA, Maisons-Alfort, France.
authors.forename: S
authors.initials: S
authors.lastname: Zientara
authors.affiliation: VISAVET Health Surveillance Centre, Universidad Complutense Madrid, Madrid, Spain.|Animal Health Department, Universidad Complutense Madrid, Madrid, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: Sánchez-Vizcaíno
mesh_terms: D000355:African Horse Sickness / Q000175:diagnosis*; D000356:African Horse Sickness Virus / Q000276:immunology; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology; D006736:Horses; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D015164:Vaccines, Inactivated; D014758:Viral Core Proteins / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000276:immunology; D014765:Viral Vaccines
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; C115108:NS3 protein, African horse sickness virus; D015164:Vaccines, Inactivated; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C069264:VP7 major core antigen, African horsesickness virus
keywords: African horse sickness; DIVA diagnosis; NS3; VP7; antibody detection; movement restriction