Improvement of an enzyme-linked immunosorbent assay for equine herpesvirus type 4 by using a synthetic-peptide 24-mer repeat sequence of glycoprotein G as an antigen.
Abstract: To increase the sensitivity of an enzyme-linked immunosorbent assay (ELISA) for equine herpesvirus type 4 (EHV-4) that uses a 12-mer peptide of glycoprotein G (gG4-12-mer: MKNNPIYSEGSL) [4], we used a longer peptide consisting of a 24-mer repeat sequence (gG4-24-mer: MKNNPIYSEGSLMLNVQHDDSIHT) as an antigen. Sera of horses experimentally infected with EHV-4 reacted much more strongly to the gG4-24-mer peptide than to the gG4-12-mer peptide. We used peptide ELISAs to test paired sera from horses naturally infected with EHV-4 (n=40). gG4-24-mer ELISA detected 37 positive samples (92.5%), whereas gG4-12-mer ELISA detected only 28 (70.0%). gG4-24-mer ELISA was much more sensitive than gG4-12-mer ELISA.
Publication Date: 2015-10-01 PubMed ID: 26424485PubMed Central: PMC4785124DOI: 10.1292/jvms.15-0275Google Scholar: Lookup
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Summary
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This research paper focuses on improving sensitivity of the enzyme-linked immunosorbent assay (ELISA) test for equine herpesvirus type 4 (EHV-4) by using a longer synthetic-peptide sequence as an antigen.
Objective of the Research
- The primary objective of the study was to enhance the effectiveness of an enzyme-linked immunosorbent assay (ELISA), a test that measures immune responses in the body, specifically targeting equine herpesvirus type 4 (EHV-4). This was achieved by using a longer synthetic-peptide sequence derived from glycoprotein G of the virus, as an antigen in place of the traditionally used 12-mer peptide.
Methodology
- The researchers created a longer peptide consisting of a 24-mer repeat sequence to replace the conventional gG4-12-mer (12-mer peptide sequence) used as an antigen in the ELISA for EHV-4.
- The serum of horses that were experimentally infected with EHV-4 was tested. The response was observed to be substantially stronger to the gG4-24-mer peptide when compared to the gG4-12-mer peptide.
- Furthermore, they utilized peptide ELISAs to test paired serum from naturally EHV-4 infected horses. The samples consisted of forty sera.
Findings and Conclusion
- The findings of the study showed that the gG4-24-mer ELISA detected 37 positive samples out of 40 (achieving a 92.5% detection rate). In contrast, the gG4-12-mer ELISA detected only 28 positive samples (representing a 70.0% detection rate).
- Therefore, the researchers concluded that the gG4-24-mer ELISA was significantly more sensitive than the gG4-12-mer ELISA.
- The research indicates a potential improvement in the detection rate of EHV-4 by increasing the length of the synthetic-peptide sequence used as an antigen in the ELISA test. By doing so, the sensitivity of the test improved, enabling a more accurate diagnosis of the disease.
Cite This Article
APA
Bannai H, Nemoto M, Tsujimura K, Yamanaka T, Maeda K, Kondo T.
(2015).
Improvement of an enzyme-linked immunosorbent assay for equine herpesvirus type 4 by using a synthetic-peptide 24-mer repeat sequence of glycoprotein G as an antigen.
J Vet Med Sci, 78(2), 309-311.
https://doi.org/10.1292/jvms.15-0275 Publication
Researcher Affiliations
- Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.
MeSH Terms
- Amino Acid Sequence
- Animals
- Antigens, Viral / immunology
- Chemistry Techniques, Synthetic
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Herpesvirus 4, Equid / isolation & purification
- Horse Diseases / virology
- Horses
- Peptide Fragments / chemical synthesis
- Peptide Fragments / immunology
- Sensitivity and Specificity
- Viral Envelope Proteins / chemical synthesis
- Viral Envelope Proteins / immunology
References
This article includes 7 references
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Citations
This article has been cited 4 times.- Bannai H, Kambayashi Y, Tsujimura K, Nagashima T, Takebe N, Tominari M, Nemoto M, Ohta M. Persistence of virus-neutralizing antibodies in horses inoculated with two doses of a live equine herpesvirus type 1 vaccine with different vaccination intervals. J Equine Sci 2021;32(3):99-102.
- Bannai H, Tsujimura K, Nemoto M, Ohta M, Yamanaka T, Kokado H, Matsumura T. Epizootiological investigation of equine herpesvirus type 1 infection among Japanese racehorses before and after the replacement of an inactivated vaccine with a modified live vaccine. BMC Vet Res 2019 Aug 6;15(1):280.
- Bannai H, Kambayashi Y, Kume K, Takebe N, Endo Y, Kawanishi N, Nemoto M, Tsujimura K. Reduction in endemic equine herpesvirus type-1 and type-4 infection among Thoroughbred yearlings through an updated vaccination program. J Equine Sci 2025 Jun;36(2):67-74.
- Fukumoto N, Bannai H, Kawanishi N, Shibata M, Kishi D, Kambayashi Y, Tsujimura K, Nemoto M. The first outbreak of equine coronavirus infection in 13 years among draft horses at Obihiro Racecourse in Japan in 2025. J Vet Med Sci 2025 Oct 1;87(10):1158-1163.
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