Validation of an indirect ELISA employing a chimeric recombinant gag and env peptide for the serological diagnosis of equine infectious anemia.
Abstract: The National Reference Center for equine infectious anemia (EIA) validated a commercial ELISA (Eradikit EIAV Indirect ELISA, In3diagnostic, Turin, Italy) employing a chimeric recombinant gag and env peptide for the detection of EIA virus antibodies, following the guidelines of the World Organization for Animal Health. The validation parameters evaluated were: analytical sensitivity (Se) and specificity (Sp); diagnostic Se and Sp; precision, based on repeatability and reproducibility through the estimation of the standard deviation (SD) and the coefficient of variation (CV); accuracy, estimated from a multiple K and relative Sp and Se with respect to those of the agar gel immunodiffusion test (AGIDT). Positive and negative predictive values were also defined. The assay showed a high specificity and a limit of detection of 1.43 log major than AGIDT. Diagnostic Se was 100% and Sp was 99.3%, while SD values ranged from 1.58 to 5.01 with a CV between 2.8% and 28.8%. Multiple K was 0.98 and relative Se and Sp were respectively 99.1% and 100%. The assay proved to be robust and to possess a high sensitivity in detecting first antibodies produced at onset of infection as well as high analytic and diagnostic Se and Sp values, confirming it as a serological assay fit for purpose within EIA surveillance programs.
Copyright © 2017 Elsevier B.V. All rights reserved.
Publication Date: 2017-10-03 PubMed ID: 28986292DOI: 10.1016/j.jviromet.2017.10.002Google Scholar: Lookup
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- Journal Article
- Validation Study
- Antibodies
- Diagnostic Technique
- Disease control
- Disease Diagnosis
- Disease Management
- Disease Outbreaks
- Disease Prevalence
- Disease Surveillance
- Disease Treatment
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Health
- Equine Infectious Anemia
- Horses
- Infectious Disease
- Serology
- Veterinary Medicine
- Veterinary Research
- Veterinary Science
- Virus
Summary
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The research article discusses the validation of an ELISA assay protocol for detecting equine infectious anemia (EIA) antibodies that utilizes a chimeric recombinant gag and env peptide. The assay demonstrates high sensitivity and specificity in identifying the first set of antibodies produced at the onset of the infection.
Overview of the Assay Validation
- The ELISA (Eradikit EIAV Indirect ELISA) was validated at the National Reference Center for equine infectious anemia (EIA) following the guidelines stipulated by the World Organization for Animal Health.
- The validation process examined several parameters such as analytical sensitivity and specificity, diagnostic sensitivity and specificity, precision, and accuracy.
- To estimate precision, standard deviation (SD) and the coefficient of variation (CV) were calculated, while a multiple K along with relative sensitivity and specificity was used to determine accuracy.
- Additionally, positive and negative predictive values for the assay were defined.
Analysis of Assay Validation Findings
- The assay demonstrated precise detection with a limit of 1.43 log major than the Agar Gel Immunodiffusion Test (AGIDT).
- It exhibited a high diagnostic sensitivity of 100% and specificity of 99.3%, thus indicating its ability to accurately detect positive cases and correctly identify negative ones.
- The standard deviation values ranged from 1.58 to 5.01 with a coefficient of variation between 2.8% and 28.8%, reflecting a reliable degree of precision.
Interpretation and Conclusion
- A multiple K value of 0.98 and relative sensitivity and specificity values of 99.1% and 100% respectively, demonstrate the accuracy of the ELISA.
- The assay displayed a high level of robustness and sensitivity, indicating its ability to identify the initial antibodies produced at the start of an EIA infection.
- Overall, the research validates the ELISA as a strong serological diagnostic tool to be used in EIA surveillance programs due to its high analytical and diagnostic sensitivity and specificity.
Cite This Article
APA
(2017).
Validation of an indirect ELISA employing a chimeric recombinant gag and env peptide for the serological diagnosis of equine infectious anemia.
J Virol Methods, 251, 111-117.
https://doi.org/10.1016/j.jviromet.2017.10.002 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Antibodies, Viral / blood
- Antigens, Viral / genetics
- Antigens, Viral / immunology
- Enzyme-Linked Immunosorbent Assay / methods
- Equine Infectious Anemia / diagnosis
- Gene Products, env / genetics
- Gene Products, env / immunology
- Gene Products, gag / genetics
- Gene Products, gag / immunology
- Horses
- Infectious Anemia Virus, Equine / immunology
- Predictive Value of Tests
- Recombinant Proteins / genetics
- Recombinant Proteins / immunology
- Reproducibility of Results
- Sensitivity and Specificity
- Serologic Tests / methods
Grant Funding
- 001 / World Health Organization
Citations
This article has been cited 2 times.- Malossi CD, Fioratti EG, Cardoso JF, Magro AJ, Kroon EG, Aguiar DM, Borges AMCM, Nogueira MF, Ullmann LS, Araujo JP Jr. High Genomic Variability in Equine Infectious Anemia Virus Obtained from Naturally Infected Horses in Pantanal, Brazil: An Endemic Region Case.. Viruses 2020 Feb 12;12(2).
- de Pablo-Maiso L, Doménech A, Echeverría I, Gómez-Arrebola C, de Andrés D, Rosati S, Gómez-Lucia E, Reina R. Prospects in Innate Immune Responses as Potential Control Strategies against Non-Primate Lentiviruses.. Viruses 2018 Aug 17;10(8).
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