Establishment of an enzyme-linked immunosorbent assay for Getah virus infection in horses using a 20-mer synthetic peptide for the E2 glycoprotein as an antigen.
Abstract: An enzyme-linked immunosorbent assay (ELISA) using a synthetic peptide for the E2 glycoprotein was developed for the serodiagnosis of Getah virus infection in horses. To identify an immunogenic epitope, a series of 20-mer peptides (n = 22) for the E2 protein was screened with pooled sera from horses infected with Getah virus. Peptide P11 (PTEEEIDMHTPPDIPDITLL) showed the strongest reaction. ELISA using P11 (E2-P11-ELISA) detected increased antibody levels in all seven experimentally infected horses and in five out of nine vaccinated horses. Out of 28 naturally infected horses, 25 were seronegative in their acute sera but turned seropositive in their convalescent sera. For the remaining three horses whose acute sera were seropositive, an endpoint method with serial dilutions detected a ≥ 4-fold increase in titer between paired sera. The concordance between E2-P11-ELISA and a virus-neutralization test in terms of seropositivity was assessed using a series of 220 horse sera, resulting in almost perfect agreement, with a kappa coefficient value of 0.865. E2-P11-ELISA had a sensitivity of 93.3% (95% CI 86.6-97.1%) and a specificity of 95.0% (95% CI 92.5-96.4%). This highly sensitive and specific E2-P11-ELISA should be useful for serodiagnosis of Getah virus infection in horses.
Publication Date: 2019-12-18 PubMed ID: 31853643DOI: 10.1007/s00705-019-04508-2Google Scholar: Lookup
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Summary
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The study developed a successful diagnostic tool (known as enzyme-linked immunosorbent assay, or ELISA) for Getah virus infection in horses using a specific peptide for the E2 glycoprotein. The research confirmed that the diagnostic tool had a sensitivity of 93.3% and a specificity of 95%, and therefore can be used effectively to detect this equine disease.
Development of ELISA Using Synthetic Peptide for the E2 Glycoprotein
- The researchers sought to develop an ELISA for diagnosing Getah virus infection in horses. This virus is a significant concern in horse management.
- The team used a synthetic peptide for the E2 glycoprotein in the virus. A synthetic peptide is a short sequence of amino acids designed to mimic a specific part of a protein, in this case, the E2 glycoprotein.
- Through various tests, they identified one peptide (P11) that showed the strongest reaction to the virus, making it an effective antigen for the test. An antigen is a substance that triggers an immune response in the body, crucial for detecting a virus.
ELISA Test Effectiveness
- The newly developed E2-P11-ELISA was effective in detecting increased antibody levels in all seven experimentally infected horses and in five out of nine vaccinated horses. This shows its potential reliability in different scenarios of infection and immunity.
- It was also effective in diagnosing natural infections with 25 out of 28 horses testing negative in their acute phase and positive in their convalescent phase. The remaining three horses, already positive in the acute phase, showed a significant rise in antibody levels on serological testing, further signifying the test’s usefulness under different stages of disease progression.
Comparison of ELISA and Virus-Neutralization Test
- Researchers compared the performance of the E2-P11-ELISA to a virus-neutralization test, a standard procedure for diagnosing viral infections. Over a series of 220 horse sera, the match between these two tests was found to be almost perfect, as evidenced by a kappa coefficient of 0.865, a statistical measure of agreement between two diagnostic tests.
- The newly developed E2-P11-ELISA showed a sensitivity of 93.3% and a specificity of 95.0%, indicating a high ability to correctly identify both infected and non-infected horses.
- The high sensitivity and specificity values of this newly developed ELISA suggest that it can be a robust and reliable diagnostic tool for Getah virus infection in horses.
Cite This Article
APA
Bannai H, Nemoto M, Tsujimura K, Ohta M.
(2019).
Establishment of an enzyme-linked immunosorbent assay for Getah virus infection in horses using a 20-mer synthetic peptide for the E2 glycoprotein as an antigen.
Arch Virol, 165(2), 377-385.
https://doi.org/10.1007/s00705-019-04508-2 Publication
Researcher Affiliations
- Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi, 329-0412, Japan. hiroshi_bannai@jra.go.jp.
- Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
- Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
- Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi, 329-0412, Japan.
MeSH Terms
- Alphavirus / genetics
- Alphavirus Infections / diagnosis
- Alphavirus Infections / veterinary
- Alphavirus Infections / virology
- Amino Acid Sequence
- Animals
- Antibodies, Viral / genetics
- Antibodies, Viral / immunology
- Antigens, Viral / genetics
- Antigens, Viral / immunology
- Enzyme-Linked Immunosorbent Assay / methods
- Glycoproteins / genetics
- Glycoproteins / immunology
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses / virology
- Peptides / genetics
- Peptides / immunology
- Sensitivity and Specificity
- Serologic Tests / methods
Citations
This article has been cited 5 times.- Qiu X, Cao X, Shi N, Zhang H, Zhu X, Gao Y, Mai Z, Jin N, Lu H. Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein. Front Microbiol 2022;13:1029444.
- Sun Q, Xie Y, Guan Z, Zhang Y, Li Y, Yang Y, Zhang J, Li Z, Qiu Y, Li B, Liu K, Shao D, Wang J, Ma Z, Wei J, Li P. Seroprevalence of Getah virus in Pigs in Eastern China Determined with a Recombinant E2 Protein-Based Indirect ELISA. Viruses 2022 Sep 30;14(10).
- Zhao J, Zhang R, Zhu L, Deng H, Li F, Xu L, Huan J, Sun X, Xu Z. Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein. BMC Vet Res 2021 Nov 19;17(1):355.
- Lan J, Duan L, Liu X, Zhou Y, Zeng B, Chen S, Ye Y, Huang D, Wan G, Zhang F, Song D. Seroprevalence of Getah virus in pigs in Southeast China determined with a recombinant Cap protein-based indirect ELISA. Front Microbiol 2025;16:1547670.
- Zhong D, Zheng J, Ma Z, Wang Y, Wei J. Rapid Detection of Getah Virus Antibodies in Horses Using a Recombinant E2 Protein-Based Immunochromatographic Strip. Animals (Basel) 2024 Aug 8;14(16).
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