Development and evaluation of a Sarcocystis neurona-specific IgM capture enzyme-linked immunosorbent assay.
Abstract: Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease of horses caused primarily by the protozoal parasite Sarcocystis neurona. Currently available antemortem diagnostic testing has low specificity. The hypothesis of this study was that serum and cerebrospinal fluid (CSF) of horses experimentally challenged with S neurona would have an increased S neurona-specific IgM (Sn-IgM) concentration after infection, as determined by an IgM capture enzyme linked immunoassay (ELISA). The ELISA was based on the S neurona low molecular weight protein SNUCD-1 antigen and the monoclonal antibody 2G5 labeled with horseradish peroxidase. The test was evaluated using serum and CSF from 12 horses experimentally infected with 1.5 million S neurona sporocysts and 16 horses experimentally infected with varying doses (100 to 100,000) of S neurona sporocysts, for which results of histopathologic examination of the central nervous system were available. For horses challenged with 1.5 million sporocysts, there was a significant increase in serum Sn-IgM concentrations compared with values before infection at weeks 2-6 after inoculation (P < .0001). For horses inoculated with lower doses of S neurona, there were significant increases in serum Sn-IgM concentration at various points in time after inoculation, depending on the challenge dose (P < .01). In addition, there was a significant increase between the CSF Sn-IgM concentrations before and after inoculation (P < .0001). These results support further evaluation of the assay as a diagnostic test during the acute phase of EPM.
Publication Date: 2006-04-06 PubMed ID: 16594589DOI: 10.1892/0891-6640(2006)20[322:daeoas]2.0.co;2Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Cerebrospinal Fluid
- Clinical Pathology
- Diagnosis
- Diagnostic Technique
- Disease
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Health
- Equine Protozoal Myeloencephalitis
- Experimental Methods
- Horses
- Immunoglobulin M
- Immunology
- Infection
- Monoclonal Antibodies
- Neurological Diseases
- Protozoa
- Sarcocystis
- Serum
- Veterinary Medicine
- Veterinary Research
Summary
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The research investigates a specific ELISA test for detecting Sarcocystis neurona, a parasite causing a serious neurological disease in horses known as EPM. The article demonstrates how this test could improve the specificity of current diagnostic methods.
Objective and Hypothesis of the Research
- The goal of the study was to enhance the specificity of current diagnostic testing for Equine protozoal myeloencephalitis (EPM). This disease is predominantly caused by the protozoal parasite Sarcocystis neurona.
- The researchers speculated that horses infected with S neurona would show increased levels of S neurona-specific IgM (Sn-IgM) in their serum and cerebrospinal fluid. This increase could be tracked via an IgM capture enzyme-linked immunoassay (ELISA).
Procedure of the Experiment
- The ELISA was predicated on the S neurona low molecular weight protein SNUCD-1 antigen and the monoclonal antibody 2G5.
- The test was carried out on serum and cerebrospinal fluid (CSF) from 28 horses. A group of 12 horses were infected with 1.5 million S neurona sporocysts. The other 16 horses were also infected with S neurona sporocysts at varied doses from 100 to 100,000 sporocysts, and their central nervous system was available for histopathologic examination.
Results of the Experiment
- For horses infected with 1.5 million sporocysts, there was a significant increase in serum Sn-IgM concentrations 2-6 weeks after inoculation.
- In the case of horses inoculated with lower doses of S neurona, significant increases in serum Sn-IgM concentration were noticed at different times post-inoculation, based on the dose they were challenged with.
- Additionally, a significant upsurge was seen in the CSF Sn-IgM concentrations before and after inoculation.
Conclusion
- The research indicates that the IgM capture ELISA test based on S neurona can potentially improve the diagnosis of EPM during its acute phase.
- Further evaluation of the assay is encouraged to ascertain its effectiveness as a diagnostic test.
Cite This Article
APA
Murphy JE, Marsh AE, Reed SM, Meadows C, Bolten K, Saville WJ.
(2006).
Development and evaluation of a Sarcocystis neurona-specific IgM capture enzyme-linked immunosorbent assay.
J Vet Intern Med, 20(2), 322-328.
https://doi.org/10.1892/0891-6640(2006)20[322:daeoas]2.0.co;2 Publication
Researcher Affiliations
- Department of' Clinical Sciences, The Ohio State University, Columbus, USA.
MeSH Terms
- Animals
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Horse Diseases / blood
- Horse Diseases / cerebrospinal fluid
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Immunoglobulin M / analysis
- Immunoglobulin M / blood
- Immunoglobulin M / cerebrospinal fluid
- Immunoglobulin M / immunology
- Sarcocystis / immunology
- Sarcocystosis / blood
- Sarcocystosis / cerebrospinal fluid
- Sarcocystosis / diagnosis
- Sarcocystosis / veterinary
- Sensitivity and Specificity
- Time Factors
Citations
This article has been cited 2 times.- Zoll WM, Prakoso D, Dark M, Liu J, Stockdale-Walden H, Long MT. Histologic characterization of eosinophilic encephalitis in horses in Florida. J Vet Diagn Invest 2018 May;30(3):442-446.
- Dubey JP, Howe DK, Furr M, Saville WJ, Marsh AE, Reed SM, Grigg ME. An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM). Vet Parasitol 2015 Apr 15;209(1-2):1-42.
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