Validation of a serum ELISA test for cyathostomin infection in equines

Kirsty L Lightbody, Andrew Austin, Peter A Lambert, Georg von Samson-Himmelstjerna, Laura Jürgenschellert, Jürgen Krücken, Martin K Nielsen, Guillaume Sallé, Fabrice Reigner, Callum G Donnelly, Carrie J Finno, Nicola Walshe, Grace Mulcahy, Nicola Housby-Skeggs, Steven Grice, Kathrin K Geyer, Corrine J Austin, Jacqueline B Matthews

Research output: Contribution to journalArticlepeer-review


Cyathostomins are ubiquitous equine nematodes. Infection can result in larval cyathostominosis due to mass larval emergence. Although faecal egg count (FEC) tests provide estimates of egg shedding, these correlate poorly with burden and provide no information on mucosal/luminal larvae. Previous studies describe a serum IgG(T)-based ELISA (CT3) that exhibits utility for detection of mucosal/luminal cyathostomins. Here, this ELISA is optimised/validated for commercial application using sera from horses for which burden data were available. Optimisation included addition of total IgG-based calibrators to provide standard curves for quantification of antigen-specific IgG(T) used to generate a CT3-specific 'serum score' for each horse. Validation dataset results were then used to assess the optimised test's performance and select serum score cut-off values for diagnosis of burdens above 1,000, 5,000 and 10,000 cyathostomins. The test demonstrated excellent performance (Receiver Operating Characteristic Area Under the Curve values >0.9) in diagnosing infection, with >90% sensitivity and >70% specificity at the selected serum score cut-off values. CT3-specific serum IgG(T) profiles in equines in different settings were assessed to provide information for commercial test use. These studies demonstrated maternal transfer of CT3-specific IgG(T) in colostrum to newborns, levels of which declined before increasing as foals consumed contaminated pasture. Studies in geographically distinct populations demonstrated that the proportion of horses that reported as test positive at a 14.37 CT3 serum score (1,000-cyathostomin threshold) was associated with parasite transmission risk. Based on the results, inclusion criteria for commercial use were developed. Logistic regression models were developed to predict probabilities that burdens of individuals are above defined thresholds based on the reported serum score. The models performed at a similar level to the serum score cut-off approach. In conclusion, the CT3 test provides an option for veterinarians to obtain evidence of low cyathostomin burdens that do not require anthelmintic treatment and to support diagnosis of infection.

Original languageEnglish
Pages (from-to)23-32
Number of pages10
JournalInternational journal for parasitology
Issue number1
Early online date1 Aug 2023
Publication statusPublished - Jan 2024

Bibliographical note

Copyright © 2023 The Author(s). Published by Elsevier Ltd on behalf of Australian Society for Parasitology. This is an open access article under the CC BY-NC-ND license (

Funding Information:The technical transfer component of these studies was supported by the Horse Trust, Bucks, UK (Registered Charity:
231748, Grant number: G3017)


  • Cyathostomin
  • Diagnostic
  • Horse
  • Nematode
  • Small strongyles


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