Development of an Indirect ELISA to Detect Equine Antibodies to Theileria haneyi.
Abstract: The apicomplexan parasite is one of two known causative agents of equine theileriosis. It causes milder clinical disease than its more virulent counterpart, , in experimentally infected horses, and can superinfect -positive horses. The current equi merozoite antigen 1 (EMA1)-based competitive enzyme-linked immunosorbent assay (ELISA)used in the U.S. to detect equine theileriosis detects but not , and the complexity of molecular assays precludes widespread use for epidemiologic studies. In order to facilitate urgently needed studies on the prevalence of , the goal of this study was to develop a sensitive and specific serologic assay for the diagnosis of based on the equi merozoite antigen 11 (EMA11). To achieve this objective, EMA11 was recombinantly expressed in eukaryotic cells and its antigenicity assessed using sera from -experimentally infected horses. Confirmation of sera reactivity enabled design and optimization of an indirect ELISA. Specificity of the ELISA for was assessed using a cohort of sera from horses experimentally infected and confirmed PCR-positive for either or . Data from field samples further demonstrate that the EMA11 ELISA is capable of identifying antibodies in horses from multiple continents around the world.
Publication Date: 2021-02-27 PubMed ID: 33673478PubMed Central: PMC7997436DOI: 10.3390/pathogens10030270Google Scholar: Lookup
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- Journal Article
- Antibodies
- Diagnosis
- Diagnostic Technique
- Disease
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Diseases
- Equine Health
- Equine Science
- Horses
- Immunology
- Infection
- Infectious Disease
- Laboratory Methods
- Parasites
- Serodiagnosis
- Theileria equi
- Veterinary Care
- Veterinary Medicine
- Veterinary Research
Summary
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The new research article focuses on the development of an Indirect ELISA to effectively detect equine antibodies of Theileria haneyi, a less virulent agent causing equine theileriosis, a disease in horses.
Understanding Theileria Haneyi and its Detection
- The research is geared towards the detection and study of theileriosis in horses. Theileriosis is a disease caused by the apicomplexan parasites Theileria equi and Theileria haneyi. This study focuses on T. haneyi, which results in a milder clinical disease compared to T. equi.
- Currently used diagnostic tools in the U.S., such as ELISA based on the equi merozoite antigen 1 (EMA1), are capable of detecting T. equi but not T. haneyi.
- Additionally, complex molecular assays used to detect these equine parasites are not consistent and widely usable for widespread epidemiological studies.
EMA11-based Diagnostic Approach
- The study aimed to develop a sensitive and precise diagnostic tool based on the equi merozoite antigen 11 (EMA11) to determine the prevalence of T. haneyi in horses around the world.
- This was accomplished by recombinantly expressing EMA11 in eukaryotic cells, followed by analyzing its antigenicity by using sera from T. haneyi-experimentally infected horses.
- After confirming the reactivity of the serum, an indirect Enzyme-Linked Immunosorbent Assay (ELISA) was designed and optimized. ELISA is a common laboratory test used in the detection and quantification of antibodies in the blood, signifying the presence of infection.
Evaluation of ELISA Assay
- Next, the specificity of the newly developed EMA11 ELISA was evaluated by using a set of sera from horses that had been experimentally infected and PCR-confirmed to either T. haneyi or T. equi.
- The study found that the EMA11 ELISA was able to pinpoint the T. haneyi antibodies effectively. This was concluded by comparing the reactions of both sets of sera and confirming that the EMA11 ELISA was specific to T. haneyi.
- Field samples were also used to further ascertain the ELISA’s able to identify T. haneyi antibodies in horses from multiple continents worldwide, indicating its usefulness for global epidemiological studies of T. haneyi.
Cite This Article
APA
Bastos RG, Sears KP, Dinkel KD, Kappmeyer L, Ueti MW, Knowles DP, Fry LM.
(2021).
Development of an Indirect ELISA to Detect Equine Antibodies to Theileria haneyi.
Pathogens, 10(3), 270.
https://doi.org/10.3390/pathogens10030270 Publication
Researcher Affiliations
- Department of Veterinary Microbiology & Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
- Department of Veterinary Microbiology & Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
- Department of Veterinary Microbiology & Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
- Animal Disease Research Unit, USDA-ARS, Pullman, WA 99164, USA.
- Department of Veterinary Microbiology & Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
- Animal Disease Research Unit, USDA-ARS, Pullman, WA 99164, USA.
- Department of Veterinary Microbiology & Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
- Department of Veterinary Microbiology & Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
- Animal Disease Research Unit, USDA-ARS, Pullman, WA 99164, USA.
Grant Funding
- #2090-320000-039-00D / Agricultural Research Service
Conflict of Interest Statement
The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.
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This article includes 20 references
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Citations
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