Improved Serodiagnosis of Hydatid Cyst Disease Using Gold Nanoparticle Labeled Antigen B in Naturally Infected Sheep
نویسندگان
چکیده
BACKGROUND Cystic echinococcosis caused by the metacestode of Echinococcus granulosus is a major problem in both humans and domestic animals health. Therefore, a standardized and approachable diagnostic tool (rapid tests) for the serodiagnosis of cystic echinococcosis (CE) is still needed. METHODS In the present work, antigen B labeled with gold nanoparticles was used to detect antibodies against hydatid cyst disease. The prepared antigen B was analyzed by SDS-PAGE. Tetra chloroauric acid (HAuCl4) was used to produce colloidal gold and antigen B labeled by gold nanoparticles, then it was tested by using rabbits antisera and sera from naturally infected sheep. The labeled antigen B was evaluated using Dot-immunogold staining (Dot-IGS) method. RESULTS Electrophoretic pattern of hydatid cyst fluid showed the quality of bands in the condensed fluid is better than crude fluid. SDS-PAGE analysis cyst fluid and antigen B revealed three specific protein bands that were detected at molecular weights of 24, 30 and 40 kDa that all are the subunits of antigen B. Evaluation of antigen B labeled by gold nanoparticles by using Dot-IGS technique showed 1/1 and 1/50 dilutions in comparison with another has the best immunoreaction. In this method, nanoparticles produced a typical purple color, when they binded to the strip at the site of immunoreaction. CONCLUSION Therefore, using gold nanoparticles is a good candidate for detection of helminthiasis, also as selective tools of early detection, simple and cost-effective, regardless of specific skills and equipment with optimal durability.
منابع مشابه
Evaluation of Enzyme Linked Immunosorbent Assay, Utilizing Native Antigen B for Serodiagnosis of Human Hydatidosis
Background: Hydatidosis is one of the cosmopolitan parasitic zoonoses caused by the larval stage of Echinococcus granulosus. Diagnosis of hydatidosis is still an unresolved problem. Serological tests using crude antigens for diagnosis of E. granulosus are sensi-tive, however their specificity are not satisfactory. Therefore, WHO recommended spe-cific serological methods using specific antigens,...
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Hydatid cyst fluid has been used as a source of antigen for the serodiagnosis of Echinococcus granulosus infection. Due to cross-reactions with antigens shared by other helminthes, the specific antigen from hydatid cyst fluid was purified by many workers. We used a relatively simple technique for purification of specific antigen from sheep hydatid cyst fluid. Isopycnic ultracentrifugation w...
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BACKGROUND Hydatidosis is one of the cosmopolitan parasitic zoonoses caused by the larval stage of Echinococcus granulosus. Diagnosis of hydatidosis is still an unresolved problem. Serological tests using crude antigens for diagnosis of E. granulosus are sensitive, however their specificity are not satisfactory. Therefore, WHO recommended specific serological methods using specific antigens, sp...
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