Hostname: page-component-848d4c4894-xm8r8 Total loading time: 0 Render date: 2024-06-15T14:22:39.822Z Has data issue: false hasContentIssue false

Copro-antigen capture ELISA for the detection of Teladorsagia (Ostertagia) circumcincta in sheep: improvement of specificity by heat treatment

Published online by Cambridge University Press:  10 June 2004

D. A. JOHNSON
Affiliation:
School of Biology, University of Nottingham, University Park, Nottingham NG7 2RD, UK
J. M. BEHNKE
Affiliation:
School of Biology, University of Nottingham, University Park, Nottingham NG7 2RD, UK
G. C. COLES
Affiliation:
Department of Clinical Veterinary Science, University of Bristol, Langford House, Bristol BS40 5DU, UK

Abstract

A copro-antigen capture ELISA for the detection of Teladorsagia circumcincta infection in sheep was developed and evaluated. Experiments with faeces from worm-free sheep, that had been spiked with known concentrations of excretory–secretory (E–S) antigen indicated that a positive signal was generated with 180 ng of E–S/ml. A nested design, based on 8 infected and 8 worm-free sheep, was employed to assess the stages during sample preparation contributing to variation in signal from the assay. This showed that 87% of the variance in the optical density readings (ODs) was directly explained by infection status. Variation between individual sheep within infection groups, and between samples at various stages in the assay, collectively accounted for the remaining variance. Initial evaluation of specificity using faeces from animals carrying a range of monospecific infections indicated cross-reactivity with Haemonchus contortus and Nematodirus spathiger. However, by treating the supernatant from faeces for 20 min at 100 °C, the cross-reactive signal was eliminated whilst the specific signal was largely preserved. Heat treatment of faeces from 12 non-infected sheep, 12 sheep with T. circumcincta and 6 with H. contortus resulted in sensitivity being increased from 66·7 to 85·7%, and specificity from 62·5 to 87·5%. OD values showed a significant positive relationship with adult worm burdens, although at low infection intensities there was some overlap between infected and worm-free animals. We discuss the application of CC-ELISAs in facilitating selective chemotherapy of sheep, as a means of avoiding the development of anthelmintic resistance in pastoral regions where sheep are farmed on a large scale.

Type
Research Article
Copyright
© 2004 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

AGNEESSENS, J., CLAEREBOUT, E. & VERCRUYSSE, J. (2001). Development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle. Veterinary Parasitology 97, 227238.CrossRefGoogle Scholar
ALLAN, J. C., CRAIG, P. S., GARCIA NOVAL, J., MENCOS, F., LIU, D., WANG, Y., WEN, H., ZHOU, P., STRINGER, R., ROGAN, M. & ZEYHLE, E. (1992). Coproantigen detection for immunodiagnosis of echinococcosis and taeniasis in dogs and humans. Parasitology 104, 347355.CrossRefGoogle Scholar
ALLAN, J. C., MENCOS, F., GARCIA NOVAL, J., SARTI, E., FLISSER, A., WANG, Y., LIU, D. & CRAIG, P. S. (1993). Dipstick dot ELISA for the detection of Taenia coproantigens in humans. Parasitology 107, 7985.CrossRefGoogle Scholar
ALLAN, J. C., VELASQUEZ-TOHOM, M., TORRES-ALVAREZ, R., YURRITA, P. & GARCIA-NOVAL, J. (1996). Field trial of the coproantigen based diagnosis of Taenia solium taeniasis by enzyme linked immunosorbent assay. American Journal of Tropical Health and Hygiene 54, 352356.CrossRefGoogle Scholar
BAUMANN, D. & GOTTSTEIN, B. (1987). A double antibody sandwich ELISA for the detection of Entamoeba histolytica in stool samples of humans. Tropical Medicine and Parasitology 38, 8185.Google Scholar
BRADFORD, M. M. (1976). A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principle of protein-dye binding. Analytical Biochemistry 72, 248254.CrossRefGoogle Scholar
COLES, G. C. (1995). Chemotherapy of human nematodes: learning from the problems of sheep. Journal of the Royal Society of Medicine 88, 649P651P.Google Scholar
CONDOR, G. A. & CAMPBELL, W. C. (1995). Chemotherapy of nematode infections of veterinary importance, with special reference to drug resistance. Advances in Parasitology 35, 184.CrossRefGoogle Scholar
DEPLAZES, P., GOTTSTEIN, B., ECKERT, J., JENKINS, D. J., EWALD, D. & JIMINEZ-PALACIOS, S. (1992). Detection of Echinococcus coproantigens by enzyme-linked immunosorbent assay in dogs, dingoes and foxes. Parasitology Research 78, 303308.CrossRefGoogle Scholar
ELLIS, T. M., GREGORY, A., TURNOR, R., KALKHOVEN, M. & WROTH, R. H. (1993). Detection of Haemonchus contortus surface antigen in faeces from infected sheep. Veterinary Parasitology 51, 8597.CrossRefGoogle Scholar
GALEN, R. S. & GAMBINO, S. R. (1975). Beyond Normality: The Predictive Value of Medical Diagnosis. Wiley, New York.
GREEN, E. L. (1986). Immunological detection of parasite antigen in faeces. Parasitology Today 2, 198200.CrossRefGoogle Scholar
GREEN, E. L., WARHURST, D. C. & MILES, M. A. (1985). Immunodiagnostic detection of Giardia antigen in faeces by a rapid visual enzyme linked immunosorbent assay. The Lancet 2, 691694.CrossRefGoogle Scholar
HENNESSY, D. R. (1997). Livestock parasite treatment – a call for greater interaction between research and industry sectors. International Journal for Parasitology 27, 11291133.CrossRefGoogle Scholar
JACKSON, F. & COOP, R. L. (2000). The development of anthelmintic resistance in sheep nematodes. Parasitology 120, S95S107.CrossRefGoogle Scholar
JACKSON, F., JACKSON, E. & COOP, R. L. (1992). Evidence of multiple anthelminthic resistance in a strain of Teladorsagia circumcincta (Ostertagia circumcincta) isolated from goats in Scotland. Research in Veterinary Science 53, 371374.CrossRefGoogle Scholar
JELENIK, T., PEYERL, G., LOSCHER, T. H. & NOTHDURFT, H. D. (1996). Giardiasis in travellers: evaluation of an antigen-capture ELISA for the detection of Giardia lamblia-antigen in stool. Zeitschrift für Gastroenterologie 34, 237240.Google Scholar
JOHNSON, D. A. (1999). Coproantigen capture ELISA for the detection of intestinal nematode infections. Ph.D. thesis. School of Biological Sciences, University of Nottingham.
JOHNSON, D. A., BEHNKE, J. M. & COLES, G. C. (1999). Development of a coproantigen capture ELISA for the detection of Teladorsagia circumcincta in sheep. In Proceedings of the 17th WAAVP Conference, Copenhagen, Denmark.
JOHNSON, M. (1994). Coproantigen ELISA for GI nematodes. Ph.D. thesis. School of Biological Sciences, University of Nottingham.
JOHNSON, M. J., BEHNKE, J. M. & COLES, G. C. (1995). Detection of gastrointestinal nematodes by a coproantigen capture ELISA. Research in Veterinary Science 60, 712.Google Scholar
NAGESWAREN, C., CRAIG, P. S. & DEVANEY, E. (1994). Coproantigen detection in rats experimentally infected with Strongyloides ratti. Parasitology 108, 335342.CrossRefGoogle Scholar
NICHOLLS, J. & OBENDORF, D. L. (1994). Application of a composite faecal egg count procedure in diagnostic parasitology. Veterinary Parasitology 52, 337342.CrossRefGoogle Scholar
RODRIGUEZ-CANUL, R., FRASER, A., ALLAN, J. C., DOMINGUEZ-ALPIZAR, J. L., ARGAEZ-RODRIGUEZ, F. & CRAIG, P. S. (1999). Epidemiological study of Taenia solium taeniasis/cysticercosis in a rural village in Yucatan state, Mexico. Annals of Tropical Medicine and Parasitology 93, 5767.CrossRefGoogle Scholar
ROEPSTORFF, A. (1998). Natural Ascaris suum infections in swine diagnosed by coprological and serological (ELISA) methods. Parasitology Research 84, 537543.CrossRefGoogle Scholar
SMITH, G. (1990). Chemotherapy: future problems. In Hookworm Disease. Current Status and New Directions (ed. Schad, G. A. & Warren, K. S.), pp. 291303. Taylor & Francis, London.
STEAR, M. J., BISHOP, S. C., DOLIGALSKA, M., DUNCAN, J. L., HOLMES, P. H., IRVINE, J., McCRIRIE, L., McKELLAR, Q. A., SINSKI, E. & MURRAY, M. (1995). Regulation of egg production, worm burden, worm length and worm fecundity by host responses in sheep infected with Ostertagia circumcincta. Parasite Immunology 17, 643652.CrossRefGoogle Scholar
STEAR, M. J. & MURRAY, M. (1994). Genetic resistance to parasitic disease: particularly of resistance in ruminants to gastrointestinal nematodes. Veterinary Parasitology 54, 161176.CrossRefGoogle Scholar
STRAIN, S. A. J., BISHOP, S. C., HENDERSON, N. G., KERR, A., McKELLAR, Q. A., MITCHELL, S. & STEAR, M. J. (2002). The genetic control of IgA activity against Teladorsagia circumcincta and its association with parasite resistance in naturally infected sheep. Parasitology 124, 545552.CrossRefGoogle Scholar
URQUHART, G. M., ARMOUR, J., DUNCAN, J. L., DUNN, A. M. & JENNINGS, F. W. (1987). Veterinary Parasitology. Avon: Longman Scientific & Technical.
VAN WYK, J. A. (1990). Occurrence and dissemination of anthelmintic resistance in South Africa, and management of resistant worm strains. In Resistance of Parasites to Antiparasitic Drugs (ed. Boray, J. C., Martin, P. J. & Roush, R. T.), pp. 103113. MSD, AGVET, Rahway, New Jersey, USA.
WALLER, P. J. (1986). Anthelmintic resistance in nematode parasites of sheep. Agricultural Zoology Reviews 1, 333373.Google Scholar
WALLER, P. J. (1993). Control strategies to prevent resistance. Veterinary Parasitology 46, 133142.CrossRefGoogle Scholar
WALLER, P. J., DASH, K. M., BARGER, I. A., LE JAMBRE, L. F. & PLANT, J. (1995). Anthelminthic resistance in nematode parasites of sheep: learning from the Australian experience. Veterinary Record 136, 411414.CrossRefGoogle Scholar