Comparison of two commercial kits and an in-house ELISA for the detection of equine rotavirus in foal feces.
Abstract: Group A rotaviruses (RVA) are important infectious agents associated with diarrhea in the young of several animal species including foals. Currently, a variety of diagnosis methods are commercially available, like ELISA, latex agglutination and immunochromatographic assays. These commercial tests are mainly designed for the detection of human RVA; its applicability in veterinary diagnosis has been poorly studied. The aim of this study was to compare the sensitivity and specificity of two commercial diagnostic kits, Pathfinder™ Rotavirus and FASTest Rota® strip, with an in-house KERI ELISA, for the detection of equine RVA. A total of 172 stool samples from Thoroughbred foals with diarrhea were analyzed. The presence of equine RVA in samples in which only one of the three methods showed positive results was confirmed by RT-PCR. A sample was considered "true positive" when RVA was detected by at least two of the methods, and "true negative" when it tested negative by the three assays. Following these criteria, 50 samples were found positive and 122 were found negative, and were handled as reference population for the assay validation. Pathfinder™ Rotavirus assay showed 32% sensitivity and 97% specificity, FASTest Rota® strip, 92% sensitivity and 97% specificity, and KERI ELISA, 76% sensitivity and 93% specificity. Pathfinder™ Rotavirus showed 77%, FASTest Rota® strip 95%, and KERI ELISA 88% accuracy to correctly classify the samples as equine RVA positive or negative. Pathfinder failed specifically to detect equine RVA G3P12I6 genotype; such performance might be related to the specificity of the monoclonal antibody included in this kit. According to our results, differences among VP6 genotypes could influence the sensitivity to detect equine RVA in foal feces, and thus assay validation of diagnostic kits for each species is necessary. In conclusion, FASTest Rota® strip is more suitable than ELISA Pathfinder™ Rotavirus for the screening of rotavirus infection in foals. The KERI ELISA showed an acceptable performance, and could be considered a proper economic alternative for equine RVA diagnosis.
Copyright © 2015 Elsevier B.V. All rights reserved.
Publication Date: 2015-05-13 PubMed ID: 25979610DOI: 10.1016/j.jviromet.2015.05.002Google Scholar: Lookup
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- Comparative Study
- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Clinical Study
- Comparative Study
- Diagnosis
- Diagnostic Technique
- Diarrhea
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Health
- Foals
- Genotyping
- Horses
- In Vivo
- Infection
- Infectious Disease
- Laboratory Methods
- Monoclonal Antibodies
- Polymerase Chain Reaction
- Rotavirus
- Sensitivity and Specificity
- Veterinary Medicine
Summary
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The research article describes a study comparing the efficiency of two commercial kits and an in-house ELISA for detection of equine rotavirus in foal feces.
Objective of the Research
- The primary aim of this research was to examine and contrast the sensitivity and specificity of two commercial diagnostic kits – Pathfinder™ Rotavirus and FASTest Rota® strip – with an in-house KERI ELISA, a diagnostic test used to detect rotavirus in equine.
Research Methodology
- A total of 172 stool samples from Thoroughbred foals experiencing diarrhea were analyzed.
- The presence of equine rotavirus in samples showing positive results in only one of the three testing methods was confirmed by RT-PCR, a laboratory technique used for detecting the virus.
- Samples that tested positive for rotavirus in at least two of the three tests were categorized as “true positive”, and those testing negative in all assays were deemed “true negative”.
- Fifty samples were found positive and 122 were found negative based on these criteria, forming the reference population for the assay validation.
Results
- The results revealed 32% sensitivity and 97% specificity for Pathfinder™ Rotavirus assay, 92% sensitivity, and 97% specificity for FASTest Rota® strip, and 76% sensitivity and 93% specificity for KERI ELISA.
- Pathfinder™ Rotavirus demonstrated 77% accuracy, FASTest Rota® strip showed 95% accuracy, and KERI ELISA recorded 88% accuracy in correctly categorizing the samples as equine rotavirus positive or negative.
- Pathfinder notably failed to detect a specific genotype of rotavirus (equine RVA G3P12I6), which could be due to the monoclonal antibody specificity included in the kit.
Conclusion
- FASTest Rota® strip outperformed the ELISA Pathfinder™ Rotavirus in screening for rotavirus infection in foals, based on the study’s outcomes.
- The KERI ELISA, however, exhibited good performance and could be an affordable alternative for equine rotavirus diagnosis.
- The study indicates that variations among VP6 genotypes could impact sensitivity when detecting equine rotavirus, thereby underscoring the importance of validating each diagnostic kit for specific species.
Cite This Article
APA
Miño S, Kern A, Barrandeguy M, Parreño V.
(2015).
Comparison of two commercial kits and an in-house ELISA for the detection of equine rotavirus in foal feces.
J Virol Methods, 222, 1-10.
https://doi.org/10.1016/j.jviromet.2015.05.002 Publication
Researcher Affiliations
- Institutode Virología, CICVyA, INTA-Castelar, Nicolás Repetto y De los Reseros s/n (1686), Hurlingham Buenos Aires, Argentina.
- MEGACOR Diagnostk GmbH Lochauer Str. 2 A 6912 Hörbranz, Austria.
- Institutode Virología, CICVyA, INTA-Castelar, Nicolás Repetto y De los Reseros s/n (1686), Hurlingham Buenos Aires, Argentina; Escuela de Veterinaria, Universidad del Salvador, Champagnat 1599, Ruta Panamericana km54.5 (B1630AHU), Pilar, Buenos Aires, Argentina.
- Institutode Virología, CICVyA, INTA-Castelar, Nicolás Repetto y De los Reseros s/n (1686), Hurlingham Buenos Aires, Argentina. Electronic address: vparreno@cnia.inta.gov.ar.
MeSH Terms
- Animals
- Diagnostic Tests, Routine / methods
- Feces / virology
- Horses
- Immunoassay / methods
- Molecular Sequence Data
- RNA, Viral / genetics
- Rotavirus / isolation & purification
- Rotavirus Infections / veterinary
- Rotavirus Infections / virology
- Sensitivity and Specificity
- Sequence Analysis, DNA
- Veterinary Medicine / methods
Citations
This article has been cited 5 times.- Nemoto M, Matsumura T. Equine rotavirus infection. J Equine Sci 2021 Mar;32(1):1-9.
- Kim JS, Lee SK, Ko DH, Hyun J, Kim HS. Performance Evaluation of the Automated Fluorescent Immunoassay System Rotavirus Assay in Clinical Samples. Ann Lab Med 2019 Jan;39(1):50-57.
- Soltan MA, Tsai YL, Lee PA, Tsai CF, Chang HG, Wang HT, Wilkes RP. Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species. J Virol Methods 2016 Sep;235:99-104.
- Chen Y, Wu J, Gao EB, Lu Y, Qiu H. A rapid visualization method for detecting rotavirus A by combining nuclear acid sequence-based amplification with the CRISPR-Cas12a assay. J Med Microbiol 2024 Oct;73(10).
- Carossino M, Vissani MA, Barrandeguy ME, Balasuriya UBR, Parreño V. Equine Rotavirus A under the One Health Lens: Potential Impacts on Public Health. Viruses 2024 Jan 16;16(1).
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