Evaluation of the specificity of five oligoprobes for identification of cyathostomin species from horses.
Abstract: Here, we report evaluation of five oligoprobes designed from intergenic spacer (IGS) region sequences for identification of cyathostomin species. Oligoprobes were designed for identification of Cylicocyclus ashworthi, Cylicocyclus nassatus, Cylicostephanus longibursatus, Cylicostephanus goldi and a fifth probe designed to identify all members of this tribe. PCR amplification of IGS DNA from 16 cyathostomin species allowed sequence comparison and identification of four putative species-specific probes. Southern blotting of amplified products from 16 species showed that all probes were species-specific. The fifth probe recognised all 16 cyathostomin species but did not bind to members of the genus Strongylus. Furthermore, these probes were used to identify individual infective L3, eggs and L4 indicating that they will be invaluable to furthering the study of the epidemiology and pathogenesis of these important equine nematodes.
Publication Date: 2001-03-10 PubMed ID: 11239940DOI: 10.1016/s0020-7519(00)00161-2Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article presents the evaluation of five oligoprobes developed for identifying cyathostomin species in horses using the intergenic spacer (IGS) sequence region.
Objective and Methodology
- The primary objective of this research was to evaluate five oligoprobes designed for the identification of specific cyathostomin species. Cyathostomins are a significant internal parasitic threat to horses. Recognizing species-specific differences in these parasites can support efforts to diagnose and manage equine parasitic diseases.
- The oligoprobes were designed based on sequences from the intergenic spacer (IGS) region for identification of the four species: Cylicocyclus ashworthi, Cylicocyclus nassatus, Cylicostephanus longibursatus, Cylicostephanus goldi and a fifth one was able to recognize all members of this tribe.
- The method adopted for evaluation involved polymerase chain reaction (PCR) amplification of IGS DNA from 16 cyathostomin species. Sequences were compared and potential species-specific probes were identified. Southern blotting, a technique for detecting specific DNA sequences in DNA samples, was used on amplified products from the 16 species.
Findings of the Study
- The results from the study confirmed that all the oligoprobes were species-specific. This means each oligoprobe was only binding to the DNA of the specific cyathostomin species it was designed to identify.
- The fifth oligoprobe was capable of recognizing all 16 cyathostomin species but did not bind to members of a different nematode genus, Strongylus.
- Moreover, these probes were used to identify individual infective L3, eggs, and L4 stages of the parasites, providing evidence that they can be used for studying the life-cycle, epidemiology, and pathogenesis of these equine nematodes.
Significance of the Research
- This research is invaluable for advancing our understanding of cyathostomin species. The use of these highly specific oligoprobes enhances the precision in studying the interaction between parasitic nematodes and their equine hosts. This, in turn, can inform better methods of diagnosis and treatment.
- Apart from the immediate context, this study also demonstrates the general utility of designing species-specific oligoprobes using the IGS region.
Cite This Article
APA
Hodgkinson JE, Love S, Lichtenfels JR, Palfreman S, Ramsey YH, Matthews JB.
(2001).
Evaluation of the specificity of five oligoprobes for identification of cyathostomin species from horses.
Int J Parasitol, 31(2), 197-204.
https://doi.org/10.1016/s0020-7519(00)00161-2 Publication
Researcher Affiliations
- Division of Equine Studies, Department of Veterinary Clinical Sciences and Animal Husbandry, University of Liverpool, Leahurst, South, CH64 7TE, Wirral, UK. jhodgkin@liverpool.ac.uk
MeSH Terms
- Animals
- Base Sequence
- DNA, Helminth / analysis
- DNA, Helminth / genetics
- DNA, Ribosomal Spacer / analysis
- DNA, Ribosomal Spacer / genetics
- Horse Diseases / parasitology
- Horses
- Molecular Sequence Data
- Oligonucleotide Probes
- Polymerase Chain Reaction / methods
- Sequence Alignment
- Sequence Analysis, DNA
- Species Specificity
- Strongylida Infections / parasitology
- Strongylida Infections / veterinary
- Strongyloidea / classification
- Strongyloidea / genetics
- Strongyloidea / isolation & purification
Citations
This article has been cited 6 times.- Johnson ACB, Biddle AS. The Use of Molecular Profiling to Track Equine Reinfection Rates of Cyathostomin Species Following Anthelmintic Administration. Animals (Basel) 2021 May 9;11(5).
- Nabavi R, Conneely B, McCarthy E, Good B, Shayan P, DE Waal T. Comparison of internal transcribed spacers and intergenic spacer regions of five common Iranian sheep bursate nematodes. Iran J Parasitol 2014 Sep;9(3):350-7.
- Traversa D, Otranto D. Biotechnological advances in the diagnosis of little-known parasitoses of pets. Parasitol Res 2009 Jan;104(2):209-16.
- Traversa D, Iorio R, Klei TR, Kharchenko VA, Gawor J, Otranto D, Sparagano OA. New method for simultaneous species-specific identification of equine strongyles (nematoda, strongylida) by reverse line blot hybridization. J Clin Microbiol 2007 Sep;45(9):2937-42.
- Hodgkinson JE, Freeman KL, Lichtenfels JR, Palfreman S, Love S, Matthews JB. Identification of strongyle eggs from anthelmintic-treated horses using a PCR-ELISA based on intergenic DNA sequences. Parasitol Res 2005 Mar;95(4):287-92.
- Wang T, Chen X, Yan X, Su Y, Gao W, Liu C, Wang W. Progress in serology and molecular biology of equine parasite diagnosis: sustainable control strategies. Front Vet Sci 2025;12:1663577.
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