Detection of Opisthorchis Viverrini by Monoclonal Antibody-Based ELISA and DNA Hybridization

Stitaya Sirisinha Faculty of Science, Mahidol University, Bangkok, Thailand

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Runglawan Chawengkirttikul Faculty of Science, Mahidol University, Bangkok, Thailand

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Rasana Sermswan Faculty of Science, Mahidol University, Bangkok, Thailand

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Sorujsiri Amornpant Faculty of Science, Mahidol University, Bangkok, Thailand

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Skorn Mongkolsuk Faculty of Science, Mahidol University, Bangkok, Thailand

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Sakol Panyim Faculty of Science, Mahidol University, Bangkok, Thailand

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Monoclonal antibody-based enzyme-linked immunosorbent assay and DNA hybridization techniques were developed and evaluated for their potential in the detection of Opisthorchis viverrini infection in humans. A mixture of three IgG1 monoclonal antibodies (MAb) specific for the 89 kDa metabolic product of O. viverrini was captured on a microtiter plate by rabbit anti-mouse IgG and used in a sandwich ELISA for the detection of parasite antigen. The 89 kD component bound to the MAb was detected with biotinylated rabbit IgG antibody to O. viverrini metabolic products. As little as 0.05–0.1 ng of the antigen could be detected by this technique. A specific O. viverrini DNA probe constructed from a repetitive DNA segment containing 340 base pairs was used in a dot blot hybridization for the detection of parasite DNA. The labeled probe constructed as such could detect DNA released from as few as five O. viverrini eggs. Both methods were specific for O. viverrini and their sensitivity was comparable with that of the classical parasitological technic.

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