Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-11-08T03:09:43.096Z Has data issue: false hasContentIssue false

Dot immunogold filtration assay (DIGFA) for the rapid detection of specific antibodies against the rat lungworm Angiostrongylus cantonensis (Nematoda: Metastrongyloidea) using purified 31-kDa antigen

Published online by Cambridge University Press:  28 May 2013

P. Eamsobhana*
Affiliation:
Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok10700, Thailand
X.X. Gan
Affiliation:
Institute of Parasitic Diseases, Zhejiang Academy of Medical Sciences, Hangzhou310013, PR China
A. Ma
Affiliation:
Institute of Parasitic Diseases, Zhejiang Academy of Medical Sciences, Hangzhou310013, PR China
Y. Wang
Affiliation:
Institute of Parasitic Diseases, Zhejiang Academy of Medical Sciences, Hangzhou310013, PR China
D. Wanachiwanawin
Affiliation:
Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok10700, Thailand
H.S. Yong
Affiliation:
Institute of Biological Sciences, University of Malaya, 50603Kuala Lumpur, Malaysia

Abstract

A rapid dot immunogold filtration assay (DIGFA) was adopted for specific immunodiagnosis of human cerebral angiostrongyliasis, using purified 31-kDa glycoprotein specific to Angiostrongylus cantonensis as diagnostic antigen and protein A colloidal gold conjugate as antigen–antibody detector. A total of 59 serum samples were assayed – 11 samples from clinically diagnosed patients with detectable A. cantonensis-specific antibody in immunoblotting; 23 samples from patients with other related parasitic diseases, i.e. gnathostomiasis (n= 8), cysticercosis (n= 5), toxocariasis (n= 2), filariasis (n= 4), paragonimiasis (n= 2) and malaria (n= 2); and 25 samples from normal healthy subjects. The sensitivity and specificity of DIGFA to detect anti-A. cantonensis specific antibodies in serologically confirmed angiostrongyliasis cases, were both 100%. No positive DIGFA was observed in cases with other parasitic diseases, and the healthy control subjects. The 3-min DIGFA is as sensitive and specific as the 3-h immunoblot test in angiostrongyliasis confirmed cases that revealed a 31-kDa reactive band. The gold-based DIGFA is more rapid and easier to perform than the traditional enzyme-linked immunosorbent assay (ELISA). The test utilizing purified A. cantonensis antigen is reliable and reproducible for specific immunodiagnosis of human infection with A. cantonensis – thus can be applied as an additional routine test for clinical diagnostic support. Large-scale sero-epidemiological studies in endemic communities in north-east Thailand are under way to evaluate its usefulness under field conditions.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2013 

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

Ali, I.O., Sibel, E., Salih, E., Haluk, O. & Hadi, A. (2005) Diagnostic value of a dot immunobinding assay for human pulmonary hydatidosis. Korean Journal of Parasitology 43, 1518.Google Scholar
Brada, D. & Roth, J. (1984) Golden blot-detection of polyclonal and monoclonal antibodies bound to antigens on nitrocellulose by protein A–gold complexes. Analytical Biochemistry 142, 79.CrossRefGoogle ScholarPubMed
Chandler, J., Gurmin, T. & Robinson, N. (2000) The place of gold in rapid tests. IVD Technology 6, 3749.Google Scholar
Chun, P.K. & Chu, A.E. (1989) A simplified 5 min staining procedure for HIV Western blots using Protein-A colloidal gold. International Conference AIDS 5, 307.Google Scholar
Cross, J.H. & Chen, E.R. (2007) Angiostrongyliasis. pp. 263292in Black, S.J. & Seed, J.R. (Eds) Food-borne parasitic zoonoses. New York, Springer Science.Google Scholar
Eamsobhana, P. (1994) Immunological studies on the rat lung-worm Angiostrongylus cantonensis (Nematoda: Metastrongylidae). PhD Thesis, University of Malaya.Google Scholar
Eamsobhana, P. (2006) The rat lungworm Angiostrongylus (Parastrongylus) cantonensis: parasitology, immunology, eosinophilic meningitis, epidemiology and laboratory diagnosis. Bangkok, Wankaew (IQ) Book Center.Google Scholar
Eamsobhana, P. (2010) Towards precise and rapid diagnosis of eosinophilic meningitis due to the rat lungworm Angiostrongylus cantonensis. Siriraj Medical Journal 62, 8489.Google Scholar
Eamsobhana, P. & Yong, H.S. (2009) Immunological diagnosis of human angiostrongyliasis cantonensis. International Journal of Infectious Diseases 13, 425431.Google Scholar
Eamsobhana, P., Yoolek, A., Suvouttho, S. & Suvuttho, S. (2001) Purification of a specific immunodiagnostic Parastrongylus cantonensis antigen by electroelution from SDS-polyacrylamide gels. Southeast Asian Journal of Tropical Medicine and Public Health 32, 308313.Google Scholar
Eamsobhana, P., Yoolek, A. & Punthuprapasa, P. (2003) Dot-blot ELISA for the immunological detection of specific antibody to Parastrongylus cantonensis. Tropical Biomedicine 20, 16.Google Scholar
Eamsobhana, P., Yoolek, A., Punthuprapasa, P. & Suvouttho, S. (2004) A dot-blot ELISA comparable to immunoblot for the specific diagnosis of human parastrongyliasis. Journal of Helminthology 78, 287291.CrossRefGoogle ScholarPubMed
Eliades, P., Karagouni, E., Stergiatou, I. & Miras, K. (1998) A simple method for the serodiagnosis of human hydatid disease based on a protein A/colloidal dye conjugate. Journal of Immunological Methods 218, 123132.Google Scholar
Feng, X., Wen, H., Zhang, Z., Chen, X., Ma, X., Zhang, J., Qi, X., Bradshaw, H., Vuitton, D. & Craig, P.S. (2010) Dot immunogold filtration assay (DIGFA) with multiple native antigens for rapid serodiagnosis of human cystic and alveolar echinococcosis. Acta Tropica 113, 114120.Google Scholar
Gan, X.X., Yue, W., Xun, G.J., Kui, Z.J. & Hua, S.X. (2006) Development of dot immunogold filtration assay kit for rapid detection of specific antibodies against Cysticercus cellulosae. Journal of Tropical Medicine Guangzhou 6, 1719.Google Scholar
Gan, X.X., Wang, Y., Guo, J.X., Yang, F.Z., Zhang, L.L. & Eamsobhana, P. (2007) Rapid detection of specific IgG antibody for Angiostrongylus cantonensis by dot immunogold filtration assay. Chinese Journal of Zoonoses 23, 345347.Google Scholar
Gan, X.X., Wang, Y., Yang, F.Z., Zhang, L. & Eamsobhan, P. (2010) Rapid detection of specific IgG to Angiostrongylus cantonensis by dot immunogold filtration assay. pp. 5966in Eamsobhana, P. (Ed.) Angiostrongylus and angiostrongyliasis – advances in the disease, control, diagnosis and molecular genetics. Bangkok, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University.Google Scholar
Graeff-Teixeira, C., da Silva, A.C.A. & Yoshimura, K. (2009) Update on eosinophilic meningoencephalitis and its clinical relevance. Clinical Microbiology Reviews 22, 322348.Google Scholar
Gui, W.J., Wang, S.T., Guo, Y.R. & Zhu, G.N. (2008) Development of a one-step strip for the detection of triazophos residues in environmental samples. Analytical Biochemistry 377, 202208.Google Scholar
Horisberger, M. & Clerr, M.F. (1985) Labelling of colloidal gold with protein A. Histochemistry 82, 219221.Google Scholar
Huang, Q., Lan, X.P., Tong, T., Wu, X., Chen, M., Feng, X.G., Liu, R.C., Tang, Y.C. & Zhu, Z.Y. (1996) Dot-immunogold filtration assay as a screening test for syphilis. Journal of Clinical Microbiology 34, 20112013.Google Scholar
Huang, Q., Tang, Y.C., Liu, W.P., Yu, S.X., Lan, X.P., Xu, B., Wu, Y.S., Li, L. & Zhu, Z.Y. (1998) Qualitative bedside assay of increased human serum myoglobin by sandwich dot-immunogold filtration for the diagnosis of acute myocardial infarction. Clinica Chimica Acta 273, 119130.Google Scholar
Liu, D.Y., Hu, W.Q. & Zhang, H.M. (2001) Application of dot immunogold filtration assay for detecting serum antibodies in clonorchiasis patients. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 19, 9799.Google Scholar
Mansour, W.A., El-Assa, F., El-Mahdy, M.M., Hendawy, M.A., Salem, R. & Demerdash, Z.A. (2009) A monoclonal antibody-based dot immunogold filtration assay: a rapid field applicable technique for immunodiagnosis of active schistosomiasis. Australian Journal of Basic and Applied Sciences 3, 784789.Google Scholar
Shim, W.B., Yang, Z.Y., Kim, J.S., Kim, J.Y., Kang, S.J., Chung, Y.C., Eremin, S.A. & Chung, D.H. (2007) Development of immunochromatography strip-test using nanocolloidal gold-antibody probe for the rapid detection of aflatoxin B1 in grain and feed samples. Journal of Microbiology and Biotechnology 17, 16291637.Google ScholarPubMed
Spielberg, F., Ryder, R.W. & Harris, J. (1989) Field testing and comparative evaluation of rapid, visually read screening assays for antibody to human immunodeficiency virus. Lancet 333, 580584.CrossRefGoogle Scholar
Wang, Y., Shi, X.H. & Gan, X.X. (2007) Dot immuno-gold filtration assay in the diagnosis of suspected paragonimiasis and evaluation of chemotherapeutic effect. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 25, 6568.Google Scholar
Wang, Q.P., Lai, D.H., Zhu, X.Q., Chen, X.G. & Lun, Z.R. (2008) Human angiostrongyliasis. Lancet Infectious Diseases 8, 621630.Google Scholar
Wen, L.Y., Chen, J.H., Ding, J.Z., Zhang, J.F., Lu, S.H., Yu, L.L., Shen, L.Y., Wu, G.L., Zhou, X.N. & Zheng, J. (2005) Evaluation on the applied value of the dot immunogold filtration assay (DIGFA) for rapid detection of anti-Schistosoma japonicum antibody. Acta Tropica 96, 142147.Google Scholar
Wu, W., Xu, D.Z., Yan, Y.P., Zhang, J.X., Liu, Y. & Li, R.L. (1999) Evaluation of dot immunogold filtration assay for anti-HAV IgM antibody. World Journal of Gastroenterology 5, 132134.CrossRefGoogle ScholarPubMed
Xiao, X., Wang, T. & Tian, Z. (2003) Development of rapid, sensitive, dye immunoassay for schistosomosis diagnosis: a colloidal dye immuno-filtration assay. Journal of Immunological Methods 280, 4957.Google Scholar