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Comparison of commercial enzyme-linked immunosorbent assays and agar gel immunodiffusion tests for the serodiagnosis of equine infectious anemia.

Abstract: The purpose of this study was to estimate the performance characteristics (accuracy, detection limit, and precision) of commercially available enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) kits in comparison with a reference AGID kit for the detection of equine infectious anemia (EIA) antibodies in horses for regulatory use in Canada. A total of 285 positive and 315 negative samples by the reference AGID were tested blindly on 2 other AGID and 4 ELISA kits. Commercially available AGID kits for the serodiagnosis of EIA were found equivalent. The 3 ELISAs directed against antibodies to the p26 core protein also performed relatively well in comparison with the reference AGID, with excellent relative accuracy and acceptable precision. The single ELISA directed against antibodies to the gp45 trans-membrane viral protein yielded a lower relative sensitivity. The performance characteristics of the ELISAs directed against antibodies to p26 are, therefore, adequate to support the implementation of ELISA for regulatory purposes in Canada.
Publication Date: 2004-12-08 PubMed ID: 15581219PubMed Central: PMC1111355
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  • Comparative Study
  • Journal Article

Summary

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This research evaluates and compares the effectiveness of commercially available enzyme-linked immunosorbent assays (ELISA) and agar gel immunodiffusion (AGID) kits, with an official AGID kit, in detecting equine infectious anemia (EIA) antibodies in horses for regulatory use in Canada. The study concluded that ELISA kits targeted towards p26 antibodies show a suitable performance for this purpose in Canada.

Research Methodology

  • The researchers used a total of 600 samples, where 285 were positive, and 315 were negative for EIA antibodies, as determined by the reference AGID kit.
  • These samples were tested using two different AGID kits and four ELISA kits without knowing the initial test results (blinded study).
  • The results were later compared against the initial test results to measure their accuracy, detection limit, and precision.

Results and Interpretation

  • The commercial AGID kits tested in the study provided equivalent performance in the serodiagnosis of EIA, meaning they showed similar results to the reference AGID kit.
  • Three ELISA kits, directed against antibodies to the p26 core protein, also exhibited excellent relative accuracy and acceptable precision.
  • However, the ELISA kit targeted against antibodies to the gp45 trans-membrane viral protein demonstrated a lower relative sensitivity, making it somewhat less reliable in detecting EIA.

Conclusion and Recommendations

  • Given the performance exhibited during the study, the researchers concluded that the ELISAs kits directed against antibodies to p26 are suitable for detecting EIA for regulatory purposes in Canada.
  • The researchers recommended the implementation of these ELISA kits in the Canadian regulatory context due to their reliable performance.

Cite This Article

APA
Paré J, Simard C. (2004). Comparison of commercial enzyme-linked immunosorbent assays and agar gel immunodiffusion tests for the serodiagnosis of equine infectious anemia. Can J Vet Res, 68(4), 254-258.

Publication

ISSN: 0830-9000
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 68
Issue: 4
Pages: 254-258

Researcher Affiliations

Paré, Julie
  • St-Hyacinthe Laboratory, Canadian Food Inspection Agency, Saint-Hyacinthe, Q J2S 8E3. parejx@inspection.gc.ca
Simard, Carole

    MeSH Terms

    • Animals
    • Antibodies, Viral / blood
    • Enzyme-Linked Immunosorbent Assay / methods
    • Enzyme-Linked Immunosorbent Assay / veterinary
    • Equine Infectious Anemia / diagnosis
    • Horses
    • Immunodiffusion / methods
    • Immunodiffusion / veterinary
    • Infectious Anemia Virus, Equine / immunology
    • Reagent Kits, Diagnostic / veterinary
    • Sensitivity and Specificity
    • Time Factors

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    Citations

    This article has been cited 8 times.
    1. Bannai H, Kambayashi Y, Nemoto M, Yamanaka T, Tsujimura K. Comparison of 4 agar gel immunodiffusion kits for serologic detection of equine infectious anemia virus antibodies. J Vet Diagn Invest 2023 Jul;35(4):430-432.
      doi: 10.1177/10406387231171567pubmed: 37129383google scholar: lookup
    2. 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.
      doi: 10.1007/s00253-023-12504-5pubmed: 37039847google scholar: lookup
    3. Alnaeem AA, Hemida MG. Surveillance of the equine infectious anemia virus in Eastern and Central Saudi Arabia during 2014-2016. Vet World 2019 May;12(5):719-723.
    4. Higgins SN, Howden KJ, James CR, Epp T, Lohmann KL. A retrospective study of owner-requested testing as surveillance for equine infectious anemia in Canada (2009-2012). Can Vet J 2017 Dec;58(12):1294-1300.
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    5. Nardini R, Autorino GL, Issel CJ, Cook RF, Ricci I, Frontoso R, Rosone F, Scicluna MT. Evaluation of six serological ELISA kits available in Italy as screening tests for equine infectious anaemia surveillance. BMC Vet Res 2017 Apr 14;13(1):105.
      doi: 10.1186/s12917-017-1007-6pubmed: 28410613google scholar: lookup
    6. Cruz F, Fores P, Ireland J, Moreno MA, Newton R. Freedom from equine infectious anaemia virus infection in Spanish Purebred horses. Vet Rec Open 2015;2(1):e000074.
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    7. Singha H, Goyal SK, Malik P, Khurana SK, Singh RK. Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus. Indian J Virol 2013 Dec;24(3):349-56.
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    8. Ostuni A, Frontoso R, Crudele MA, Barca L, Amati M, Boni R, De Vendel J, Raimondi P, Bavoso A. Comparative Evaluation of a Multistrain Indirect ELISA Targeting Anti- p26 and gp45 Antibodies for EIAV Detection. Pathogens 2025 Jun 8;14(6).
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