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Applied microbiology and biotechnology2023; 107(10); 3305-3317; doi: 10.1007/s00253-023-12504-5

Development and evaluation of a blocking ELISA for serological diagnosis of equine infectious anemia.

Abstract: Equine infectious anemia (EIA) is an important viral disease characterized by persistent infection in equids worldwide. Most EIA cases are life-long virus carriers with low antibody reactions and without the appearance of clinical symptoms. A serological test with high sensitivity and specificity is required to detect inapparent infection. In this study, a B-cell common epitope-based blocking ELISA (bELISA) was developed using a monoclonal antibody together with the EIAV p26 protein labelled with HRP. The test has been evaluated against the standard and with field serum samples globally. This bELISA test can be completed within 75 min, and the sensitivity is higher than those of either the AGID or one commercial cELISA kit. This bELISA assay was 8-16 times more analytically sensitive than AGID, and 2 to 4 times more analytically sensitive than one cELISA kit by testing three sera from the USA, Argentina, and China, respectively. The 353 serum samples from Argentina were tested, in comparison with AGID, the diagnostic sensitivity and specificity of our bELISA assay were 100% (154/154) and 97.0% (193/199), respectively, and the accuracy of the bELISA test was 98.3%. The bELISA test developed in this study is a rapid, sensitive, specific method for the detection of EIAV infection, and could be a promising candidate for use in the monitoring of the EIA epidemic worldwide. KEY POINTS: • A universal epitope-based blocking enzyme-linked immunosorbent assay (bELISA) was developed for detection of antibodies to EIAV. • The bELISA assay can be used to test EIAV serum samples from different regions of the world including North America, South America, Europe, and Asia. • The bELISA assay was evaluated in three different international labs and showed a better performance than other commercial kits.
© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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Publication Date: 2023-04-11 PubMed ID: 37039847PubMed Central: 2168377DOI: 10.1007/s00253-023-12504-5Google Scholar: Lookup
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Summary

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The researchers developed and evaluated a new lab test, known as a blocking ELISA, for diagnosing equine infectious anemia. The study found that the new ELISA test is rapid, highly sensitive and specific for the detection of equine infectious anemia virus, making it a promising tool for tracking the spread of the disease worldwide.

Overview of Equine infectious anemia (EIA)

  • EIA is a significant viral disease characterized by long-lasting infection in equids around the world.
  • Most EIA cases are life-long carriers of virus with low antibody reactions and without any clinical symptoms.
  • A highly sensitive and specific serological test is required to detect inapparent infection.

Development of the Blocking ELISA (bELISA) Test

  • A B-cell common epitope-based blocking ELISA (also known as bELISA) was developed.
  • This was done using a monoclonal antibody combined with the EIAV p26 protein labelled with HRP (a substance that helps in detection).
  • The test can be completed within 75 minutes.
  • Its sensitivity is higher than those of either the AGID (a standard test) or a commercial cELISA kit.

Evaluation of the bELISA Test

  • The bELISA assay was 8-16 times more sensitive than AGID and 2 to 4 times more sensitive than one cELISA kit. This was proved by testing three sera from the USA, Argentina, and China.
  • In a trial of 353 serum samples from Argentina, compared to AGID, the diagnostic sensitivity and specificity of the bELISA test were 100% and 97.0%, respectively.
  • The accuracy of the bELISA test was 98.3%.

Potential Applications

  • The bELISA test could be used to detect EIAV infection rapidly and precisely.
  • The assay can be used to test EIAV serum samples from various regions of the world including North America, South America, Europe, and Asia.
  • It holds promise as a tool in monitoring the EIA epidemic worldwide.

Cite This Article

APA
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. (2023). Development and evaluation of a blocking ELISA for serological diagnosis of equine infectious anemia. Appl Microbiol Biotechnol, 107(10), 3305-3317. https://doi.org/10.1007/s00253-023-12504-5

Publication

Applied microbiology and biotechnology
ISSN: 1432-0614
NlmUniqueID: 8406612
Country: Germany
Language: English
Volume: 107
Issue: 10
Pages: 3305-3317

Researcher Affiliations

Hu, Zhe
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
  • WOAH Reference Laboratory for Equine Infectious Anemia, Harbin, China.
Guo, Kui
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Du, Cheng
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Sun, Jinhui
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
  • Northeast Agricultural University, Harbin, China.
Naletoski, Ivancho
  • Animal Production and Health Section, Joint FAO/IAEA Centre for Nuclear Applications in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Vienna, Austria.
Chu, Xiaoyu
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Lin, Yuezhi
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Wang, Xuefeng
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China.
Barrandeguy, Maria
  • Instituto Nacional de Tecnología Agropecuaria (INTA), Instituto de Virología, Pilar, Buenos Aires, Argentina.
  • Facultad de Ciencias Agrarias Y Veterinarias, Universidad del Salvador, Pilar, Buenos Aires, Argentina.
Samuel, Miño
  • Instituto Nacional de Tecnología Agropecuaria (INTA), Instituto de Virología, Pilar, Buenos Aires, Argentina.
Wang, Wen
  • Animal Health Supervision Institute of Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang, China.
Lau, Patrick Imtung
  • Equine Disease Division, Agriculture Fisheries and Conservation Department, Hong Kong, China.
Wernery, Ulrich
  • Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Raghavan, Rekha
  • Central Veterinary Research Laboratory, Dubai, United Arab Emirates.
Wang, Xiaojun
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, China. wangxiaojun@caas.cn.
  • WOAH Reference Laboratory for Equine Infectious Anemia, Harbin, China. wangxiaojun@caas.cn.

MeSH Terms

  • Horses
  • Animals
  • Equine Infectious Anemia / diagnosis
  • Antibodies, Viral
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Enzyme-Linked Immunosorbent Assay / methods
  • Serologic Tests / veterinary
  • Epitopes, B-Lymphocyte
  • Sensitivity and Specificity
  • Infectious Anemia Virus, Equine

Grant Funding

  • 2021YFD1800500 / the National Key R&D Program of China
  • 2022YFD1800200 / the National Key Research and Development Program of China
  • JJ2022TD2023 / the Nature Science Foundation of Heilongjiang Provience, China

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