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Immunodiagnostic Properties of Wucheraria bancrofti SXP-1, a Potential Filarial Diagnostic Candidate Expressed in Tobacco Plant, Nicotiana tabacum

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Abstract

Transgenic tobacco plants were developed expressing WbSXP-1, a diagnostic antigen isolated from the cDNA library of L3 stage larvae of Wucheraria bancrofti. This antigen produced by recombinant Escherichia coli has been demonstrated by to be successful as potential diagnostic candidate against lymphatic filariasis. A rapid format simple and qualitative flow through immune-filtration diagnostic kit has been developed for the identification of IgG antibodies to the recombinant WbSXP-1 and is being marketed by M/S Span Diagnostics Ltd in India and Africa. Here, we present the results of experiments on the transformation and expression of the same filarial antigen, WbSXP-1, in tobacco plant, Nicotiana tabacum, to produce plant-based diagnostic antigen. It was possible to successfully transform the tobacco plant with WbSXP-1, the integration of the parasite-specific gene in plants was confirmed by PCR amplification and the expression of the filarial protein by Western blotting. The immunoreactivity of the plant-produced WbSXP-1 was assessed based on its reaction with the monoclonal antibodies developed against the E. coli-produced protein. Immunological screening using clinical sera from patients indicates that the plant-produced protein is comparable to E. coli-produced diagnostic antigen. The result demonstrated that plants can be used as suitable expression systems for the production of diagnostic proteins against lymphatic filariasis, a neglected tropical infectious disease which has a negative impact on socioeconomic development. This is the first report of the integration, expression and efficacy of a diagnostic candidate of lymphatic filariasis in plants.

Key Message

Transgenic tobacco plants with WbSXP-1, a filarial diagnostic candidate, were developed. The plant-produced protein showed immunoreactivity on par with the E. coli product.

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Acknowledgments

This research was funded by a grant (LS-30/2010) from WOS-A section of Department of Science & Technology, Government of India to Mathangi Ganapathy. The authors would like to thank The Director, Centre for Biotechnology, Anna University, Chennai for allowing access to the lab and the infrastructure. Thanks are also due to Dr. N. Subramoniam, Sugarcane Breeding Institute, Coimbatore, India for his expertise and to Mr. Jeyender Sethuraman and Mr. V. Kamalakannan for their help in statistical analysis.

Conflict of Interest

We declare that we have no conflict of interest.

Declaration

All human and animal studies have been approved by the appropriate ethics committee. All persons gave their informed consent prior to their inclusion in the study. Samples used were collected in accordance with US Department of Health and Human Services Human Experimentation Guidelines and the Department of Public Health, Chennai, Tamil Nadu, India. All of the procedures followed were in accordance with the guidelines issued by the Department of Public Health, Government of Tamil Nadu, India, for dealing with human subjects. The protocols adopted are also approved by the institutional review board at the Centre for Biotechnology, Anna University, India.

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Authors and Affiliations

  1. Centre for Biotechnology, Anna University, Chennai, 600025, India

    Mathangi Ganapathy, S. Jason Charles, S. Jaiganesh & P. Kaliraj

  2. Sugarcane Breeding Institute, Coimbatore, 641 007, Tamil Nadu, India

    M. Chakravarthi, P. Harunipriya & N. Subramonian

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  1. Mathangi Ganapathy
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  2. M. Chakravarthi
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  3. S. Jason Charles
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  4. P. Harunipriya
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  6. N. Subramonian
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  7. P. Kaliraj
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Correspondence to Mathangi Ganapathy or P. Kaliraj.

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Ganapathy, M., Chakravarthi, M., Charles, S.J. et al. Immunodiagnostic Properties of Wucheraria bancrofti SXP-1, a Potential Filarial Diagnostic Candidate Expressed in Tobacco Plant, Nicotiana tabacum . Appl Biochem Biotechnol 176, 1889–1903 (2015). https://doi.org/10.1007/s12010-015-1685-0

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