Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Development of an Indirect ELISA to Detect Equine Antibodies to Theileria haneyi

Version 1 : Received: 29 January 2021 / Approved: 1 February 2021 / Online: 1 February 2021 (09:41:24 CET)

A peer-reviewed article of this Preprint also exists.

Bastos, R.G.; Sears, K.P.; Dinkel, K.D.; Kappmeyer, L.; Ueti, M.W.; Knowles, D.P.; Fry, L.M. Development of an Indirect ELISA to Detect Equine Antibodies to Theileria haneyi . Pathogens 2021, 10, 270. Bastos, R.G.; Sears, K.P.; Dinkel, K.D.; Kappmeyer, L.; Ueti, M.W.; Knowles, D.P.; Fry, L.M. Development of an Indirect ELISA to Detect Equine Antibodies to Theileria haneyi . Pathogens 2021, 10, 270.

Abstract

The apicomplexan parasite Theileria haneyi is one of two known causative agents of equine theileriosis. It causes milder clinical disease than its more virulent counterpart, Theileria equi, in experimentally infected horses, and can superinfect T. equi-positive horses. The current EMA1-based competitive ELISA used in the U.S. to detect equine theileriosis detects T. equi but not T. haneyi, and the complexity of molecular assays precludes widespread use for epidemiologic studies. In order to facilitate urgently needed studies on the prevalence of T. haneyi, the goal of this study was to develop a sensitive and specific serologic assay for the diagnosis of T. haneyi based on the equi mero-zoite antigen 11 (ThEMA11). To achieve this objective, ThEMA11 was recombinantly expressed in eukaryotic cells and its antigenicity assessed using sera from T. haneyi-experimentally infected horses. Confirmation of sera reactiv-ity enabled design and optimization of an indirect ELISA. Specificity of the ELISA for T. haneyi was assessed using a cohort of sera from horses experimentally infected and confirmed PCR-positive for either T. equi or T. haneyi. Data from field samples further demonstrate that the ThEMA11 ELISA is capable of identifying T. haneyi antibodies in horses from multiple continents around the world.

Keywords

Equine theileriosis; Theileria haneyi; enzyme-linked immunosorbent assay (ELISA), serology

Subject

Biology and Life Sciences, Anatomy and Physiology

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