Development And Evaluation Of An Antigen Detection Test For Hydatid Cyst Disease
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Date
2018-05
Authors
Khanbabaie, Sam
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Hydatid cyst disease (HCD) is a zoonotic parasitic disease which can infect
both humans and animals. Imaging and antibody detection tests have been used to
diagnose HCD, the former cannot differentiate between hydatid cyst and other
pathologies; while the latter may not be able to differentiate between past and present
infections, and may also not be suitable for evaluating treatment efficacy. In this
regard, antigen detection tests may be able to assist in addressing the above issues.
Previously it has been reported that hydatid cyst antigens can be detected in HCD
patients using various assays. However the reported results were not satisfactory,
with variable diagnostic sensitivities (25% - 83%). Therefore, this study was aimed
at developing an antigen detection test for HCD. A panel of samples comprising
serum from HCD patients (n=35), healthy individuals (n=38) and patients with other
parasitic diseases (n=13) were used. Three types of E. granulosus antigens were
prepared i.e. hydatid cyst fluid (HCF), native antigen B (nAgB) and recombinant
antigen B (rAgB). Polyclonal antibodies against each of the antigens were produced
in rabbits, followed by purification of their IgG fractions. Next, a portion of the
purified IgGs were labelled with horseradish peroxidase (HRP). Using the three
types of IgGs and three types of HRP-conjugated antibodies, various ELISA
sandwich combinations were tested. The best ELISA combination was further
optimized and evaluated using the panel of serum samples. Another portion of the
purified IgGs were conjugated to colloidal gold nanoparticles for development of
lateral flow dipstick (LFD). The dipsticks were dotted with three different IgGs, and
LFD assay using various combinations of capture and detector antibodies were
tested.
Description
Keywords
Evaluation of an antigen detection , test for hydatid cyst disease