Development of a recombinant protein-based ELISA for diagnosis of larval cyathostomin infection



Mitchell, Mairi C, Tzelos, Thomas, Handel, Ian, Mcwilliam, Hamish EG, Hodgkinson, Jane E ORCID: 0000-0001-9152-8481, Nisbet, Alasdair J, Kharchenko, Vitaliy O, Burgess, Stewart TG and Matthews, Jacqueline B
(2016) Development of a recombinant protein-based ELISA for diagnosis of larval cyathostomin infection. PARASITOLOGY, 143 (8). pp. 1055-1066.

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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.

Item Type: Article
Uncontrolled Keywords: nematode, cyathostomin, encysted larvae, ELISA, diagnosis
Depositing User: Symplectic Admin
Date Deposited: 12 Jan 2017 14:40
Last Modified: 19 Jan 2023 07:20
DOI: 10.1017/S0031182016000627
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3005207