Development of nanocolloidal gold based immunochromatographic assay for rapid detection of transgenic vegetative insecticidal protein in genetically modified crops

被引:39
作者
Kumar, Rajesh [1 ]
Singh, Chandra K. [1 ,4 ]
Kamle, Suchitra [1 ]
Sinha, Rajeshwar P. [2 ]
Bhatnagar, Raj K. [3 ]
Kachru, Devendra N. [1 ]
机构
[1] CSIR, Indian Inst Toxicol Res, Lucknow 226001, Uttar Pradesh, India
[2] Banaras Hindu Univ, Dept Bot, Varanasi 221005, Uttar Pradesh, India
[3] Int Ctr Genet Engn & Biotechnol, New Delhi 110067, India
[4] Univ S Carolina, Sch Med, Dept Pathol Microbiol & Immunol, Columbia, SC 29209 USA
关键词
Nanocolloidal gold; Vegetative insecticidal protein; Bacillus thuringiensis; Nitrocellulose membrane; Immunochromatographic strip; Genetically modified crops; BACILLUS-THURINGIENSIS; IMMUNOASSAY; NANOPARTICLES; VIP3A; SIZE;
D O I
10.1016/j.foodchem.2010.03.086
中图分类号
O69 [应用化学];
学科分类号
081704 ;
摘要
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. (C) 2010 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1298 / 1303
页数:6
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