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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 

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