Development of a recombinant protein-based ELISA for diagnosis of larval cyathostomin infection.
Abstract: Cyathostomins are ubiquitous nematodes of horses. Once ingested, they can spend a substantial time as encysted larvae in the intestinal wall. The larvae can comprise up to 90% of the total burden, with up to several million worms reported in individuals. These stages can emerge in large numbers to cause life-threatening colitis. Direct methods for detection of encysted larval burdens in live horses do not exist. Previously, two antigen complexes were identified as promising markers for infection. A component of these, cyathostomin gut associated larval antigen-1 (Cy-GALA-1), was identified following immunoscreening of a complementary DNA library. Serum immunoglobulin G(T) (IgG(T)) responses to Cy-GALA-1 were shown to inform on larval infection. Sequence analysis of polymerase chain reaction products amplified from individual worms indicated that Cy-GALA-1 was derived from Cyathostomum pateratum. As cyathostomin infections always comprise multiple species, a diagnostic test must account for this. Here, segments of the Cy-gala gene were isolated from four common species, Cyathostomum catinatum, Cylicocyclus ashworthi, Cylicostephanus goldi and Cylicostephanus longibursatus, and the associated proteins expressed in recombinant form. The specificity and immunogenicity of each protein was confirmed. Each protein was assessed by enzyme linked immuno sorbent assay (ELISA) for its ability for informing on the presence of encysted larval infection and the level of burden.
Publication Date: 2016-05-13 PubMed ID: 27174468DOI: 10.1017/S0031182016000627Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper is about the development of a diagnostic test to detect infection by cyathostomins, a type of parasite in horses, using an enzyme-linked immunosorbent assay (ELISA) that’s based on a recombinant protein.
Research Background
- The study focuses on cyathostomins, a type of nematode that is widely prevalent in horses. These parasites, after being ingested, spend a significant amount of time as encysted larvae within the horse’s intestinal wall.
- These encysted larvae can make up 90% of a horse’s total parasite burden and if they emerge in large quantities, can lead to severe colitis which can be life-threatening.
- Previously, detecting these encysted larvae in living horses has been a challenge, with no direct methods available.
- Nonetheless, a larval antigen known as Cy-GALA-1 had been identified as a potential marker for infection. This antigen produced a response in the horse’s serum immunoglobulin G(T) (IgG(T)) that could indicate a larval infection.
Methodology
- To develop a diagnostic test that can account for infections by multiple species, gene segments of Cy-GALA (Cy-gala) were isolated from four common cyathostomin species: Cyathostomum catinatum, Cylicocyclus ashworthi, Cylicostephanus goldi, and Cylicostephanus longibursatus.
- The proteins associated with these gene segments were then expressed in recombinant form, and their specificity and immunogenicity were confirmed.
Assessment with ELISA
- The researchers used an enzyme-linked immunosorbent assay (ELISA) to evaluate each protein’s capability to identify the presence of an encysted larval infection and estimate the level of parasitic burden.
- ELISA is a plate-based assay technique used for detecting and quantifying substances such as peptides, proteins, antibodies, and hormones.
Significance of the Research
- The study is significant as successfully developing a diagnostic tool based on a recombinant protein could revolutionize the detection and treatment of cyathostomin infections.
- Earlier detection could lead to more efficient treatment, reducing instances of life-threatening colitis in horses and improving overall equine health.
Cite This Article
APA
Mitchell MC, Tzelos T, Handel I, McWilliam HE, Hodgkinson JE, Nisbet AJ, Kharchenko VO, Burgess ST, Matthews JB.
(2016).
Development of a recombinant protein-based ELISA for diagnosis of larval cyathostomin infection.
Parasitology, 143(8), 1055-1066.
https://doi.org/10.1017/S0031182016000627 Publication
Researcher Affiliations
- Moredun Research Institute,Pentlands Science Park,Edinburgh EH26 0PZ,Scotland,UK.
- Moredun Research Institute,Pentlands Science Park,Edinburgh EH26 0PZ,Scotland,UK.
- Royal Dick School of Veterinary Studies,University of Edinburgh,Edinburgh EH25 9RG,Scotland,UK.
- Moredun Research Institute,Pentlands Science Park,Edinburgh EH26 0PZ,Scotland,UK.
- Department of Infection Biology,Institute of Infection and Global Health,University of Liverpool,Liverpool L69 7ZJ,England,UK.
- Moredun Research Institute,Pentlands Science Park,Edinburgh EH26 0PZ,Scotland,UK.
- I.I. Schmalhausen Institute of Zoology NAS of Ukraine,vul B Khmelnytskogo 15,Kyiv 01030,Ukraine.
- Moredun Research Institute,Pentlands Science Park,Edinburgh EH26 0PZ,Scotland,UK.
- Moredun Research Institute,Pentlands Science Park,Edinburgh EH26 0PZ,Scotland,UK.
MeSH Terms
- Amino Acid Sequence
- Animals
- Antibody Specificity
- Antigens, Helminth / genetics
- Antigens, Helminth / immunology
- DNA, Complementary / genetics
- DNA, Helminth / genetics
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Helminth Proteins / genetics
- Helminth Proteins / immunology
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Larva
- Recombinant Proteins
- Sequence Alignment
- Strongyle Infections, Equine / diagnosis
- Strongyle Infections, Equine / parasitology
- Strongyloidea / immunology
- Strongyloidea / isolation & purification
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