title: Development of dipsticks for simultaneous detection of vip3A and cry1Ab/cry1Ac transgenic proteins.
abstract: The number of genetically modified (GM) crops being cultivated and its produce reaching market is increasing every year. The transgenes (vip3A, cry1Ab, and cry1Ac) from Bacillus thuringiensis are being used by plant breeders, apart from other transgenes for developing insect pest-resistant GM crops. It is therefore necessary to develop an easy, rapid, and reliable detection assay to discriminate GM crops and non-GM crops. Dipstick strips using colloidal gold-labeled polyclonal antibodies were developed for simultaneous detection of Vip3A and Cry1Ab/CrylAc proteins. The assay was essentially based on the sandwich format of immunoassay, which was completed within 10 min, and the results were evaluated visually. The detection limits were 50 ng/mL (50 ppb) for both CrylAc and CrylAb proteins, and 100 ng/mL (100 ppb) for Vip3A protein. The developed dipsticks are suitable for on-site simultaneous screening of GM crops bearing two proteins, which, in turn, reduce cost and time of the assay.
authors.affiliation: Banaras Hindu University, Department of Botany (Centre of Advanced Study), Varanasi-221 005, India. rajeshsbhu@yahoo.com
authors.forename: Rajesh
authors.initials: R
authors.lastname: Kumar
mesh_terms: D000906:Antibodies / Q000737:chemistry; D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D002845:Chromatography; D018556:Crops, Agricultural / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000235:genetics*; D017343:Genes, Plant; D017979:Gold Colloid / Q000737:chemistry; D006460:Hemolysin Proteins / Q000235:genetics*; D007118:Immunoassay / Q000379:methods; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D019076:Transgenes / Q000235:genetics*
chemical_list: D000906:Antibodies; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D017979:Gold Colloid; D006460:Hemolysin Proteins; D010940:Plant Proteins; D011933:Reagent Kits, Diagnostic; C447139:Vip3A protein, Bacillus thuringiensis; C030956:insecticidal crystal protein, Bacillus Thuringiensis