Hostname: page-component-586b7cd67f-t7czq Total loading time: 0 Render date: 2024-11-25T18:07:10.575Z Has data issue: false hasContentIssue false
  • English
  • Français

A simplified method for the fractionation of Gnathostoma-specific antigens for serodiagnosis of human gnathostomosis

Published online by Cambridge University Press:  05 June 2009

Siripon Tuntipopipat ,
Runglawan Chawengkirttikul  and
Stitaya Sirisinha
Siripon Tuntipopipat
Affiliation:
Department of Microbiology, Faculty of Science, Mahidol University and Laboratory of Immunology, Chulabhorn Research Institute, Bangkok, Thailand
Runglawan Chawengkirttikul
Affiliation:
Department of Microbiology, Faculty of Science, Mahidol University and Laboratory of Immunology, Chulabhorn Research Institute, Bangkok, Thailand
Stitaya Sirisinha*
Affiliation:
Department of Microbiology, Faculty of Science, Mahidol University and Laboratory of Immunology, Chulabhorn Research Institute, Bangkok, Thailand
*
*Dr. Stitaya Sirisinha, Department of Microbiology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand
Get access Rights & Permissions [Opens in a new window]

Abstract

Specific immunoreactive components present in crude somatic extract and in excretory-secretory (ES) products of Gnathostoma spinigerum advanced third-stage larvae (L3) were identified by Western blotting and their diagnostic potential evaluated by indirect ELISA. Although both crude antigen preparations were highly complex, the ES antigen gave a more satisfactory diagnostic result. Most G. spinigerum specific components present in the somatic and ES preparations had molecular weights below 29 kD and were not glycosylated, judging from the concanavalin A staining pattern. Specific diagnostic antigens were prepared by subjecting the crude preparations to SDS-PAGE. Low molecular weight components were identified and electroeluted from the gel. Excess SDS was removed by the use of an ion retardation resin. The antigens obtained by this relatively simple procedure were found to be highly specific for G. spinigerum. Sensitivity, specificity, and positive and negative predictive values for the assay using the fractionated somatic antigen were 100%.

Keywords

Gnathostoma spinigerumNematodaimmunodiagnosisgnathostomosisELISA
Type
Research Papers
Information
Journal of Helminthology , Volume 67 , Issue 4 , December 1993 , pp. 297 - 304
Copyright
Copyright © Cambridge University Press 1993

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

Dharmkrong-At, A. & Sirisinha, S. (1983) Analysis of antigens from different developmental stages of Angiostrongylus cantonensis. Southeast Asian Journal of Tropical Medicine and Public Health, 14, 154–162.Google ScholarPubMed
Dharmkrong-At, A., Migasena, S., Suntharasamai, P., Bunnag, D., Priwan, R. & Sirisinha, S. (1986) Enzyme-linked immunosorbent assay for detection of antibody to Gnathostoma antigen in patients with intermittent cutaneous migratory swelling. Journal of Clinical Microbiology, 23, 847–851.Google Scholar
Galen, R. S. (1980) Predictive values and efficiency of laboratory testing. Paediatric Clinics of North America, 27, 861–869.Google Scholar
Kapp, O. H. & Vinogradov, S. N. (1978) Removal of sodium dodecyl sulfate from proteins. Analytical Biochemistry, 91, 230–235.CrossRefGoogle ScholarPubMed
Laemmli, U. K. & Favre, M. (1973) Maturation of the head of bacteriophage T4. I. DNA packing events. Journal of Molecular Biology, 80, 575–599.Google Scholar
Lowry, O. H., Rosenbrough, 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
Maleewong, W., Morakote, N., Thamasonthi, W., Charuchinda, K., Tesana, S. & Khamboonruang, C. (1988) Serodiagnosis of human gnathostomiasis. Southeast Asian Journal of Tropical Medicine and Public Health, 19, 201–205.Google ScholarPubMed
Maleewong, W., Wongkham, C., Intapan, P., Mahaisavariya, P., Danseegaew, W., Pipigool, V. & Morakote, N. (1992) Detection of circulating parasite antigens in murine gnathostomiasis by a two-site enzyme-linked immunosorbent assay. American Journal of Tropical Medicine and Hygiene, 46, 80–84.Google Scholar
Nopparatana, C., Tapchaisri, P., Setasuban, P., Chaicumpa, W. & Dekumyoy, P. (1988) Antibody responses in human gnathostomiasis. Southeast Asian Journal of Tropical Medicine and Public Health, 19, 219–224.Google ScholarPubMed
Nopparatana, C., Setasuban, P., Chaicumpa, W. & Tapchaisri, P. (1991) Purification of Gnathostoma spinigerum specific antigen and immunodiagnosis of human gnathostomiasis. International Journal for Parasitology, 21, 677–678.Google Scholar
Soesatyo, M. H. N. E., Ratanasiriwilai, W., Suntharasamai, P. & Sirisinha, S. (1987) IgE responses in human gnathostomiasis. Transactions of the Royal Society of Tropical Medicine and Hygiene, 81, 799–801.Google Scholar
Suntharasamai, P., Desakorn, V., Migasena, S., Bunnag, D. & Harinasuta, T. (1985) ELISA for immunodiagnosis of human gnathostomiasis. Southeast Asian Journal of Tropical Medicine and Public Health, 16, 274–279.Google Scholar
Tada, I., Araki, T., Matsuda, H., Araki, K., Akahane, H. & Mimori, T. (1987) A study on immunodiagnosis of gnathostomiasis by ELISA and double diffusion with special reference to the antigenicity of Gnathostoma doloresi. Southeast Asian Journal of Tropical Medicine and Public Health, 18, 444–448.Google Scholar
Tapchaisri, P., Nopparatana, C., Chaicumpa, W. & Setasuban, P. (1991) Specific antigen of Gnathostoma spinigerum for immunodiagnosis of human gnathostomiasis. International Journal for Parasitology, 21, 315–319.Google Scholar
Towbin, H., Staehelin, T. & Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets; procedure and some applications. Proceedings of the National Academy of Science of the United States of America, 76, 4350–4354.CrossRefGoogle ScholarPubMed
Tuntipopipat, S., Chawengkirttikul, R., Witoonpanich, R., Chiemchanya, S. & Sirisinha, S. (1989) Antigens, antibodies and immune complexes in cerebrospinal fluid of patients with cerebral gnathostomiasis. Southeast Asian Journal of Tropical Medicine and Public Health, 20, 439–446.Google ScholarPubMed