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Characterization of antigens of Enterobius vermicularis (pinworm) eggs
Y B N Sanduni De Kostha1, Sisira L Pathirana1, Shiroma M Handunnetti1
1Institute of Biochemistry, Molecular Biology and Biotechnology, University of Colombo, Colombo, Sri Lanka.
Abstract:
Enterobiasis (pinworm infection) caused by Enterobius vermicularis is a common parasitic infection prevalent worldwide especially in children. Infection is diagnosed by microscopic detection of E. vermicularis eggs on perianal swabs. This study aimed to characterize the antigens of E. vermicularis eggs as a preliminary step towards identifying diagnostic targets for detection in infected individuals. The study was conducted between October 2019 and February 2020, following approval from Ethics Review Committee of the Faculty of Medicine, University of Colombo (EC-19-034). E. vermicularis eggs were harvested from perianal swabs using acetone and purified with 1× PBS (pH 7.2). A portion of eggs was used for preparing antigen slides, while the rest were sonicated and vortexed with glass beads and inoculated subcutaneously (with weekly booster doses) into a Wistar rat for developing antibodies. Blood drawing from rat was done weekly for 5 weeks. Confirmation of the presence of antibodies was done by surface immunofluorescence against eggs on the antigen slides. Protein bands were determined using SDS-PAGE assay and immunogenic antigen bands were determined by reacting with antiserum after immunoblotting. The band sizes of the proteins were determined against corresponding bands of a protein ladder. Surface immunofluorescence was positive with serum obtained from day 14 post-inoculation from the Wistar rat as well as that obtained from a person with chronic enterobiasis. The most prominent and immunogenic protein bands identified from egg antigens were 21 kDa, 66 kDa, 83 kDa, 96 kDa, 112 kDa, 121 kDa, 140 kDa and 151 kDa. Methods used in this study were effective in obtaining E. vermicularis egg antigens which were immunogenic. Furthermore, surface antigens of intact eggs reacted with antibodies developed against crushed egg antigens. These findings may pave the way for the development of effective immunodiagnostics.
Insights
Researchers identified key immunogenic protein bands from Enterobius vermicularis eggs, crucial for developing new diagnostic tools for pinworm infection (enterobiasis). These findings advance the potential for effective immunodiagnostics in detecting this common parasitic disease.
Area of Science:
- Parasitology
- Immunology
- Molecular Biology
Background:
- Enterobiasis, caused by Enterobius vermicularis, is a widespread parasitic infection, particularly in children.
- Current diagnosis relies on microscopic identification of pinworm eggs from perianal swabs.
Purpose of the Study:
- To characterize Enterobius vermicularis egg antigens.
- To identify potential diagnostic targets for enterobiasis detection.
Main Methods:
- Harvesting and purification of E. vermicularis eggs from perianal swabs.
- Inducing antibody production in Wistar rats using egg antigens.
- Analyzing protein bands via SDS-PAGE and immunoblotting to identify immunogenic antigens.
- Confirming antibody presence using surface immunofluorescence.
Main Results:
- Surface immunofluorescence confirmed antibody presence from day 14 post-inoculation in rats and in individuals with chronic enterobiasis.
- Prominent immunogenic protein bands were identified at 21, 66, 83, 96, 112, 121, 140, and 151 kDa.
- Methods effectively yielded immunogenic E. vermicularis egg antigens, with surface antigens reacting to antibodies from crushed egg antigens.
Conclusions:
- The study successfully characterized immunogenic antigens from Enterobius vermicularis eggs.
- Identified protein bands represent potential targets for developing novel immunodiagnostic methods for enterobiasis.
- These findings offer a promising pathway for improved diagnostics of pinworm infections.
