Serological diagnosis of equine infectious anemia in horses, donkeys and mules using an ELISA with a gp45 synthetic peptide as antigen.
Abstract: Equine infectious anemia (EIA) is a disease caused by a Lentivirus that is currently controlled exclusively by identification of seropositive animals. In most countries, including Brazil, the official diagnostic test for EIA is the agar gel immunodiffusion test (AGID). Although this assay has a high specificity it can produce false negative reactions or equivocal results due to weak precipitation lines, especially in samples from donkeys, mules or newly infected equids. In this pioneering study, it was used overlapping synthetic peptide pools to map and identify a consensus, widely recognised antibody epitope within env encoding the EIAV envelope proteins. A 20-mer soluble peptide encompassing this epitope (pgp45) was then synthesized and tested in an indirect ELISA test. Using a panel of 859 EIA positive and negative equid serum samples, the pgp45 ELISA had 96.1% concordance, 98.6% sensitivity and 95.6% specificity respectively, when compared to AGID. The sensitivity and specificity of the pgp45 ELISA was also >90% when tested in individual equid species including horses (Equus caballus), donkeys (Equus asinus) and mules (Equus caballus x Equus asinus). Moreover, in a horse experimentally infected with the pathogenic Wyoming EIAV strain viral-specific antibodies were detected at 10 days post-infection (dpi) whereas in AGID no specific antibody was detected until 18 days of experimental infection. This peptide can now be used as an antigen in serological tests, especially for rapid screening of large numbers of equids, where it may contribute significantly in the control of EIA, especially at sites with high populations of donkeys and mules.
Copyright © 2018 Elsevier B.V. All rights reserved.
Publication Date: 2018-12-19 PubMed ID: 30576724DOI: 10.1016/j.jviromet.2018.12.009Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Antibodies
- Antigen
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Diseases
- Equine Health
- Equine Infectious Anemia
- Horses
- Infectious Disease
- Mule
- Peptides
- Serodiagnosis
- Serological Surveys
- Seroprevalence
- Veterinary Medicine
- Veterinary Research
- Viral Diseases
Summary
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This research paper details an innovative study into a more specific and potentially quicker means of diagnosing Equine Infectious Anemia (EIA) in horses, donkeys, and mules, through the use of a particular synthetic peptide (pgp45) in an Enzyme-Linked Immunosorbent Assay (ELISA) test.
Background
- EIA is a disease in equines (horses, donkeys, mules) caused by a type of virus known as a Lentivirus.
- Currently, the EIA is identified primarily through blood tests, specifically the agar gel immunodiffusion test (AGID).
- However, AGID can produce false negatives or ambiguous results, especially in donkeys and mules or in newly infected horses, due to weak precipitation lines.
Research Methodology
- Utilizing synthetic peptide pools, the researchers aimed to identify a consensus or generally recognized antibody epitope (a portion of an antigen molecule to which an antibody attaches itself) within the EIAV envelope proteins.
- They ultimately synthesized a 20-mer soluble peptide, pgp45, which covers this epitope.
- This pgp45 was then tested in an indirect ELISA test.
Results
- The pgp45 ELISA was tested on 859 equid serum samples which included known EIA positive and negative samples for comparison with the standard AGID test.
- The concordance of ELISA test results with AGID was 96.1%, displaying a sensitivity of 98.6% and specificity of 95.6%. These are measures of the ELISA test’s true positive rate and true negative rate, respectively.
- When segregated by species — horses, donkeys, mules — the ELISA’s sensitivity and specificity both remained over 90%.
- In an experimental trial on a horse infected with the pathogenic Wyoming EIAV strain, the ELISA detected specific antibodies at 10 days post-infection, compared to AGID which didn’t detect antibodies until 18 days.
Implications
- The pgp45 ELISA offers a viable alternative for serological diagnosis of EIA, particularly when testing large equine populations or for quicker detection in early infection stages.
- This could significantly enhance EIA control measures, especially in regions with substantial donkey and mule populations.
- The faster diagnosis could potentially prevent disease spread in populations and improve animal health management.
Cite This Article
APA
Naves JHFF, Oliveira FG, Bicalho JM, Santos PS, Machado-de-Ávila RA, Chavez-Olortegui C, Leite RC, Reis JKP.
(2018).
Serological diagnosis of equine infectious anemia in horses, donkeys and mules using an ELISA with a gp45 synthetic peptide as antigen.
J Virol Methods, 266, 49-57.
https://doi.org/10.1016/j.jviromet.2018.12.009 Publication
Researcher Affiliations
- Laboratório de Retroviroses da Escola de Veterinária da Universidade Federal de Minas Gerais, Brazil. Electronic address: jhelder83@yahoo.com.br.
- Laboratório de Retroviroses da Escola de Veterinária da Universidade Federal de Minas Gerais, Brazil. Electronic address: fe_goliveira@yahoo.com.br.
- Laboratório de Retroviroses da Escola de Veterinária da Universidade Federal de Minas Gerais, Brazil. Electronic address: julianambicalho@gmail.com.
- Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, Brazil. Electronic address: paulasantos.bio@gmail.com.
- Laboratório de Fisiologia Experimental, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, Brazil. Electronic address: r_andrez@yahoo.com.
- Laboratório de Imunoquímica de Toxinas Naturais do Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais, Brazil. Electronic address: olortegi@icb.ufmg.br.
- Laboratório de Retroviroses da Escola de Veterinária da Universidade Federal de Minas Gerais, Brazil. Electronic address: romulocnpq@gmail.com.
- Laboratório de Retroviroses da Escola de Veterinária da Universidade Federal de Minas Gerais, Brazil. Electronic address: jenner@ufmg.br.
MeSH Terms
- Animals
- Antibodies, Viral / blood
- Antigens, Viral / chemistry
- Antigens, Viral / immunology
- Enzyme-Linked Immunosorbent Assay / veterinary
- Equidae / immunology
- Equidae / virology
- Equine Infectious Anemia / diagnosis
- Equine Infectious Anemia / immunology
- False Negative Reactions
- Horses / immunology
- Horses / virology
- Sensitivity and Specificity
- Serologic Tests / veterinary
- Viral Envelope Proteins / chemical synthesis
- Viral Envelope Proteins / immunology
Citations
This article has been cited 3 times.- Hu Z, Guo K, Du C, Sun J, Naletoski I, Chu X, Lin Y, Wang X, Barrandeguy M, Samuel M, Wang W, Lau PI, Wernery U, Raghavan R, Wang X. Development and evaluation of a blocking ELISA for serological diagnosis of equine infectious anemia.. Appl Microbiol Biotechnol 2023 May;107(10):3305-3317.
- Aguilar-Montes de Oca S, Montes-de-Oca-Jiménez R, Carlos Vázquez-Chagoyán J, Barbabosa-Pliego A, Eliana Rivadeneira-Barreiro P, C Zambrano-Rodríguez P. The Use of Peptides in Veterinary Serodiagnosis of Infectious Diseases: A Review.. Vet Sci 2022 Oct 12;9(10).
- Resende CF, Santos AM, Cook RF, Victor RM, Câmara RJF, Gonçalves GP, Lima JG, Maciel E Silva AG, Leite RC, Dos Reis JKP. Low transmission rates of Equine infectious anemia virus (EIAV) in foals born to seropositive feral mares inhabiting the Amazon delta region despite climatic conditions supporting high insect vector populations.. BMC Vet Res 2022 Jul 22;18(1):286.
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