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.

Bead-Based Multiplex Assay for the Simultaneous Detection of Antibodies to African Swine Fever Virus and Classical Swine Fever Virus

“…This technology is widely applied in human health for different applications, such as strain identification in infections, immune response characterization (humoral and cellular), or biomarkers identification as well as other uses (30, 31). However, less work has been carried out using this technology in the veterinary field (32–38) and there are only a few commercial kits available. Moreover, when compared to conventional ELISA, previous results have shown that xMAP formats can be more sensitive and reproducible (35).…”

Re‐parameterization of a mathematical model of African horse sickness virus using data from a systematic literature search

“…The article by Sánchez‐Matamoros et al. (2016) was considered, but not included due to uncertainty of the vaccination status of the 14 experimentally infected horses. Qualitative synthesis of the information available from papers can be found in Table S5.…”

Serological evidence of African horse sickness virus infection of donkeys in Karamoja sub-region, North-eastern Uganda

“…Disease prevention is by vaccination with a polyvalent vaccine since all AHSV serotypes are present in South Africa and in most parts of sub-Saharan Africa. Several methods are employed for the diagnosis of AHSV, including virus inoculation of cell cultures, mice inoculation (Howell, 1962), postmortem, serology and molecular assays (Costa et al, 2016;Fowler et al, 2016;de Waal et al, 2016;Sánchez-Matamoros et al, 2016;Weyer et al, 2015). AHS is not contagious, but is known to be spread by insect vectors.…”