Designing a rapid test to diagnose the ovine hydatid cyst

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Department of Parasitology, Faculty of Veterinary Medicine,University of Tehran, Tehran-Iran

Abstract

Background: Using a rapid and cost benefote test for diagnosing of hydatidosis, a zoonotic, can be beneficial as a  diagnostic. Objectives: The aim of the present study was to design and assess the performance of a dipstick method for diagnosting of cystic echinococcosis. Methods: Hydatid cyst fluid antigens and homogenized antigens of protoscolex were prepared and its electrophoretic pattern was determined by SDS-PAGE.  Afterthen, for  providing the hyperimmune serum, rabbits were injected by hydatid cyst fluid and protoscolex antigens along with complete freund adjuant and then incomplete freund adjuant. The immune sera were evaluated by dot ELISA. Dipstick was prepared based on nitrocellulose paper as solid phase and coated with antigens which were dotted in the upper surface of the nitrocellulose strip. PBS was used as negative control and rabbit sera non-infected were used as positive control.  As a negative control the lower part of the strip was coated by PBS.  Each strip was floated in the serum (1:100 dilution) for 7min, washed for 7min and floated in the second antibody 7min. Afterthen, they were washed and incubated in chromogen/substrate diaminobenzidin for 2min to show the coloured band at the site of coated antigen. Results: Our findings revealed 15 protein fractions in fluid antigens and 9 protein band in protoscolex antigens at the range of 29-130 kDa and 25-90 kDa, respectively. Meanwhile, the protoscolex antigens at in 1:10 dilution and fluid antigens at in 1:20 dilution showed the best results to diagnose hyper immune serum of 1:10 dilution. Conclusions: This  rapid Dipstick assay test  can be considered as a suitable immunodiagnostic test for hydatid cyst disease; however further investigations should be done to improve its specificity through preparing highly purified antigens.

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