Volume 14, Issue 54 (4-2007)                   RJMS 2007, 14(54): 157-166 | Back to browse issues page

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Maleki F, Sadegh Hassani S. Preparation and Purification of Mature Fasciola Hepatica Worm’s Antigens for the Diagnosis of Fascioliasis. RJMS 2007; 14 (54) :157-166
URL: http://rjms.iums.ac.ir/article-1-713-en.html
Abstract:   (8689 Views)

    Background & Aim: Fascioliasis as a zoonosis is one of the important parasitic diseases which is caused when fasciola species enter the host’s body. Approximately 2.4 million people throughout the world are infected with this parasite and 180 million people are vulnerable to it. On time diagnosis and rapid treatment of the disease can substantially prevent the harmful effects of the infection on patients. In the present study, we have tried to separate, purify and assess a useful antigen and apply it as a sensitive and specific serologic method. Material & Method: In this experiment, first healthy worms were collected and then cuticles and the other organs of them were detached, homogenized and sonicated separately. Their levels of protein were measured by Bradford method. Then, all the samples were thickened and put on chromatography gel column(G200 sephadex). Four antigenic reactions for each part were observed. By using ultrafiltration method, the protein level of the samples was adjusted to a suitable level so that the obtained antigen could be used to immunize 36 rabbits. The antibodies taken from the rabbits as well as 30 samples of human sera with fascioliasis were tested with CIEP method. In the present study, fraction 2 antigen of fasciola hepatica’s cuticles showed 90.9% sensitivity and 94.4% specificity in CIEP test. The above-mentioned antigen which was identified by SDS-PAGE method had one distinct band of 60 KD, and no other band was recognized along the gel. This antigen, then, went under experiment by ELISA method which revealed that the antibody in 26 out of 30 samples of human sera and all 36 rabbits was detectable and measurable. Results: Altogether, 60 samples out of 66 including 30 human sera and 36 rabbit sera showed positive sedimentary reaction in CIEP method whereas 62 samples did so in ELISA method. Conclusion: Therefore, it is suggested that 60 KD antigen of fasciola worm’s cuticles be used as a pure antigen in serologic tests such as ELISA and CIEP. However, the results indicate high sensitivity of ELISA method to CIEP method.

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Type of Study: Research | Subject: parasitology

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