Development of nanocolloidal gold based immunochromatographic assay for rapid detection of transgenic vegetative insecticidal protein in genetically modified crops |
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| Authors: | Rajesh Kumar Chandra K. Singh Suchitra Kamle Rajeshwar P. Sinha Raj K. Bhatnagar Devendra N. Kachru |
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| Affiliation: | 1. Indian Institute of Toxicology Research (CSIR), Post Box No. 80, M.G. Marg, Lucknow 226 001, UP, India;2. Department of Botany, Banaras Hindu University, Varanasi 221 005, UP, India;3. International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110 067, India;4. School of Medicine, Department of Pathology, Microbiology and Immunology, University of South Carolina, Columbia, SC 29209, USA |
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| Abstract: | Vegetative insecticidal protein (Vip) is now being used for transgenic expression in several crops; conferring resistance against lepidopteron pests. A rapid, single step, sensitive and specific immunochromatographic (IC) strip test for the detection of recombinant Vip-S protein in the transgenic samples was developed. Polyclonal rabbit anti-Vip-S IgG conjugated to nanocolloidal gold served as a probe to detect Vip protein in test samples. The detection limit for the developed IC strip was 100 ng/ml (100 ppb) and on addition of gold enhancer the sensitivity increased to 1 ng/ml (1 ppb) of Vip-S protein. The assay was validated with transgenic brinjal samples. The assay time was less than 10 min, suitable for rapid on-site testing. No cross-reactivity was observed with other transgenic plant proteins employed for pest and weed management, i.e. Cry1Ac, Cry1Ab, and CP4-EPSPS. This on-site test offers rapid screening for a genetically modified crops having relatively new transgene (vip) entering the global market. |
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| Keywords: | Nanocolloidal gold Vegetative insecticidal protein Bacillus thuringiensis Nitrocellulose membrane Immunochromatographic strip Genetically modified crops |
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