Authors: Abdi, J.¹; Kazemi, B.²; Mohebali, M.¹; Bandehpour, M.³; Rahimi, M. T.¹; Rokni, M. B.⁴

Source: Annals of Tropical Medicine and Parasitology, Volume 104, Number 5, July 2010 , pp. 399-407(9)

Publisher: Maney Publishing

Abstract:

A 12-kDa subunit of antigen B from Echinococcus granulosus has recently been cloned, expressed and used in diagnostic ELISA to test human sera for evidence of cystic echinococcosis. The performance of the ELISA based on the recombinant antigen (rAgB) was compared with that of similar assays based on native antigen B (nAgB) or hydatid-cyst fluid.

For the preparation of the rAgB, total RNA was extracted from Ec. granulosus protoscoleces so that antigen-B complementary DNA could be synthesised, amplified by PCR, and then cloned into the pQE30 expression vector. The recombinant plasmid was transformed in Escherichia coli and induced using isopropyl-?-D-thiogalactopyrano-side. Bacterial samples were collected, lysed and then analysed by SDS–PAGE and western blotting. The recombinant protein was purified by affinity chromatography.

Although the performance of the ELISA based on cyst fluid appeared identical to that of the assay based on the recombinant antigen (with a sensitivity, specificity, positive predictive value and negative predictive value of 96.0%, 97.0%, 97.2% and 95.5%, respectively), the corresponding results for the ELISA based on nAgB (98.6%, 100%, 100% and 98.5%) were slightly better. Despite this difference (which was not statistically significant), the comparative ease with which large quantities of the recombinant antigen could be produced make the antigen a potentially useful tool in the diagnosis of cystic echinococcosis.

Document Type: Research Article

DOI: http://dx.doi.org/10.1179/136485910X12743554760261

Affiliations: 1: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, P.O. Box 14155-6446, Tehran, Iran 2: Department of Parasitology, Shahid Beheshti University (Medical Campus), Tehran, 19857- 17443, Iran, Cellular and Molecular Biology Research Centre, Shahid Beheshti University (Medical Campus), Tehran, 19857-17443, Iran 3: Cellular and Molecular Biology Research Centre, Shahid Beheshti University (Medical Campus), Tehran, 19857-17443, Iran 4: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, P.O. Box 14155-6446, Tehran, Iran; Email: roknimoh@tums.ac.ir.

Publication date: 2010-07-01

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• In 2012 Annals of Tropical Medicine and Parasitology changed its name to Pathogens and Global Health to reflect changes and developments in the subject area. View the issues of Pathogens and Global Health available online..
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