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Immunodiagnosis of human trichinellosis using excretory-secretory (ES) antigen

Published online by Cambridge University Press:  05 June 2009

Pakpimol Mahannop ,
Wanpen Chaicumpa ,
Prasert Setasuban ,
Nimit Morakote  and
Pramuan Tapchaisri
Pakpimol Mahannop
Affiliation:
Department of Helminthology, Faculty of Public Health, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
Wanpen Chaicumpa*
Affiliation:
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
Prasert Setasuban
Affiliation:
Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
Nimit Morakote
Affiliation:
Department of Helminthology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50000, Thailand
Pramuan Tapchaisri
Affiliation:
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
*
*Author for correspondence.
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Abstract

Infective first stage larvae of Trichinella spiralis were recovered from muscles of laboratory infected mice by digesting the muscles with 1% HC1–1% pepsin and collecting the larvae by modified Baerman's method. The larvae were cultivated in a serum-free medium for 18 h. The ES antigen obtained from the culture medium was used in an enzyme-linked immunosorbent assay (ELISA) for detecting IgG antibodies to T. spiralis in serum samples collected from three groups of individuals. The individuals of the first group were parasitologically confirmed trichinellosis patients, while those of group 2 were patients with other helminthiasis and group 3 were healthy, parasite-free individuals. The specificity of the assay was 100%. The sensitivity of the test was also 100% when performed on sera of group I collected at days 57 and 120 after infection. Sera collected earlier (day 23) and those collected 700 days after infection had negligible reactivity. Thus IgG-ELISA using ES antigen of the L1 was useful not only for diagnosis but also in evaluation of cure. Western blot analysis revealed that specific antigens of T. spiralis were 94, 67, 63, and 39 kilodalton components.

Keywords

Trichinella spiralisELISAWestern blot analysisexcretory-secretory antigenspecific antigenimmunodiagnosismanNematoda
Type
Research Article
Information
Journal of Helminthology , Volume 66 , Issue 4 , December 1992 , pp. 297 - 304
Copyright
Copyright © Cambridge University Press 1992

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References

REFERENCES

Au, A. C. S., Ko, R. C., Simon, J. W., Ridell, R. J., Wong, F. W. T. & Templer, M. J. (1983) Study of acute trichinosis in Churka. Specificity of enzyme-linked immunosorbent assay for IgM and, IgE antibodies to Trichinella larval antigens in diagnosis. Transactions of the Royal Society of Tropical Medicine and Hygiene, 77, 412–415.CrossRefGoogle ScholarPubMed
Feldmeier, H., Fischer, G. & Blaumeiser, G. (1987) Kinetics of humoral response during the acute and the convalescent phase of human trichinosis. Zentralblatt für Bakteriologie, Mikrobiologie and Hygiene, series A, 264, 221–234.Google ScholarPubMed
Galen, R. S. (1980) Predictive values and efficiency of laboratory testing. Pediatric Clinics of North America, 27, 861–869.CrossRefGoogle ScholarPubMed
Gamble, H. R., Anderson, W. R., Graham, C. W. & Murrell, K. D. (1983) Diagnosis of swine trichinosis by enzyme-linked immunosorbent assay (ELISA) using an excretory-secretory antigen. Veterinary Parasitology, 13, 349–361.CrossRefGoogle ScholarPubMed
Gould, S. E. (1970) Clinical pathology: Diagnostic laboratory procedures. In: Gould, S. E. Trichinosis in Man and Animals. Springfield, Charles, C. Thomas, pp. 190–221.Google Scholar
Justus, D. E. & Morakote, N. (1981) Mast cell degranulation associated with sequestration and removal of Trichinella spiralis antigen. International Archives of Allergy and Applied Immunology, 64, 371–384.CrossRefGoogle Scholar
Kagan, T. G. & Norman, L. G. (1970) The serology of trichinosis. In: Gould, S. E. Trichinosis in Man and Animals. Springfield, Charles, C. Thomas, pp. 222–268.Google Scholar
Khamboonruang, C. & Thammasonthi, W. (1987) Enzyme-linked immunosorbent assay (ELISA) in patients infected with Trichinella spiralis. In: Kim, C. W., Ruitenberg, E. G. & Teppema, J. S. (Editors). Trichinosis. Proceeding of the Fifth International Conference on Trichinellosis Survey. Reedhooks, London, pp. 263–267.Google Scholar
Knapen, V. F., Frachimont, J. H., Ruitenberg, E. J. (1980) The reliability of the enzyme-linked immunoassay (ELISA) for the detection of swine trichinosis. In: Kim, C. W., Ruitenberg, E. G. & Teppema, J. S. (Editors). Trichinosis, Proceeding of the Fifth International Conference on Trichnellosis Survey. Reedbooks, London, pp. 399–404.Google Scholar
Knapen, V. F., Frachimont, J. H., Verdonk, A. R. & Stumpf, K. U. (1982) Detection of specific immunoglobulin (IgG, IgM, IgA, IgE) and total IgE level in human trichinosis by means of enzyme-linked immunosorbent assay (ELISA). American Journal of Tropical Medicine and Hygiene, 31, 973–976.CrossRefGoogle ScholarPubMed
KO, R. C. & Yeung, H. F. (1989) Specificity of ES antigens in detection of Trichinella spiralis antibodies in Chinese pig. Tropical Biomedicine, 6,99–111.Google Scholar
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193,265–275.CrossRefGoogle ScholarPubMed
Murrell, K. D., Anderson, W. R., Schad, G. A., Hanbury, R. D., Kazacos, K. R., Gamble, H. R. & Brown, J. (1986) Field evaluation of enzyme-linked immunosorbent assay for swine trichinosis. Efficacy of the excretory-secretory antigen. American Journal of Veterinary Research, 47, 1046–1049.Google ScholarPubMed
Morakote, N., Khamboonruang, C., Siriprasert, V., Suphawitayanukul, S., Marcanatachote, S. & Thammasonthi, W. (1991) The value of enzyme-linked immunosorbent assay (ELISA) for diagnosis of human trichinosis. Tropical Medicine and Parasitology, 42, 172–174.Google ScholarPubMed
Palmer, D. F. & Cavallaro, J. K. (1980) Some concepts of quality control in immunoserology. In: Manual of Clinical Immunology, 2nd edition (Editors Rose, N. R. & Friedman, H.). American Society for Microbiology, Washington, D. C., pp. 1078–1082.Google Scholar
Pozio, E. & Khamboonruang, C. (1989) Trichinellosis in Thailand: epidemiology and biochemical identification of the aetiological agent. Tropical Medicine and Parasitology, 40,73–74.Google Scholar
Smith, H. T. (1988) Comparison of pepsin-digestion and enzyme-linked immunosorbent assay for the diagnosis of trichinosis in swine. Canadian Journal of Veterinary Medicine Research, 52,63–66.Google ScholarPubMed
Su, X. & Prestwood, A. K. (1991) A Dot-ELISA mimicry Western blot test for the detection of swine trichinosis. Journal of Parasitology, 77, 76–82.CrossRefGoogle Scholar
Taylor, S. M., Kenny, J., Mallon, T. & Davidson, W. B. (1980) The micro-ELISA for antibodies to Trichinella spiralis. Elimination of false positive reactions by antigen fractionation and technical improvements. Zentralblatt für Veterinärmedizine, 27, 764–772.CrossRefGoogle ScholarPubMed
Towbin, H., Staehelin, T. & Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheet: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America, 76, 4350–4354.CrossRefGoogle ScholarPubMed
Zarlenga, D. S. & Gamble, S. R. (1990) Molecular cloning and expression of immunodominant 53-kDa excretory-secretory antigen from Trichinellta spiralis muscle larvae. Molecular and Biochemical Parasitology, 42, 165–174.CrossRefGoogle ScholarPubMed