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

Published online by Cambridge University Press:  13 May 2016

MAIRI C. MITCHELL ,
THOMAS TZELOS ,
IAN HANDEL ,
HAMISH E. G. MCWILLIAM ,
JANE E. HODGKINSON ,
ALASDAIR J. NISBET ,
VITALIY O. KHARCHENKO ,
STEWART T. G. BURGESS  and
JACQUELINE B. MATTHEWS
MAIRI C. MITCHELL
Affiliation:
Moredun Research Institute, Pentlands Science Park, Edinburgh EH26 0PZ, Scotland, UK
THOMAS TZELOS
Affiliation:
Moredun Research Institute, Pentlands Science Park, Edinburgh EH26 0PZ, Scotland, UK
IAN HANDEL
Affiliation:
Royal Dick School of Veterinary Studies, University of Edinburgh, Edinburgh EH25 9RG, Scotland, UK
HAMISH E. G. MCWILLIAM
Affiliation:
Moredun Research Institute, Pentlands Science Park, Edinburgh EH26 0PZ, Scotland, UK Department of Microbiology and Immunology, Doherty Institute of Infection and Immunity, The University of Melbourne, Parkville, Victoria 3010, Australia
JANE E. HODGKINSON
Affiliation:
Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool L69 7ZJ, England, UK
ALASDAIR J. NISBET
Affiliation:
Moredun Research Institute, Pentlands Science Park, Edinburgh EH26 0PZ, Scotland, UK
VITALIY O. KHARCHENKO
Affiliation:
I.I. Schmalhausen Institute of Zoology NAS of Ukraine, vul B Khmelnytskogo 15, Kyiv 01030, Ukraine
STEWART T. G. BURGESS
Affiliation:
Moredun Research Institute, Pentlands Science Park, Edinburgh EH26 0PZ, Scotland, UK
JACQUELINE B. MATTHEWS*
Affiliation:
Moredun Research Institute, Pentlands Science Park, Edinburgh EH26 0PZ, Scotland, UK
*
*Corresponding author: Moredun Research Institute, Pentlands Science Park, Edinburgh EH23 0PZ, Scotland, UK. E-mail: jacqui.matthews@moredun.ac.uk
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Summary

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.

Keywords

nematodecyathostominencysted larvaeELISAdiagnosis
Type
Research Article
Information
Parasitology , Volume 143 , Issue 8 , July 2016 , pp. 1055 - 1066
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Copyright
Copyright © Cambridge University Press 2016 

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Footnotes

†

These authors contributed equally to this work.

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