Detection of transgenic events in maize using immunochromatographic strip test and conventional PCR

Abstract

With the growth in the transgenic market, fast and economically viable methodologies are necessary for undertaking transgene detection tests, both for identification of contamination in seeds and in grain. Seeds from commercial conventional GNZ 2004, and transgenic VT-Pro (MON89034), Roundup Ready (NK603) and Herculex (TC1507) maize cultivars were used. In order to simulate different levels of contamination, the transgenic seeds were mixed with conventional seeds at levels of 0.2%, 0.4%, 1.0% and 1.6% for VT-Pro, and 0.2%, 0.5%, 0.8% and 1.2% for Roundup Ready and Herculex. The lateral flow membrane strip test was performed in the whole seed, endosperm and embryo. For evaluation of the specificity of the technique in detection of the TC1507 event by means of the conventional PCR technique, seeds of the commercial maize hybrid GNZ 2004 were used as negative control, and the maize hybrid 2B655Hx as positive control. In order to simulate different levels of contamination, transgenic seeds were mixed with conventional seeds at the levels of 10%, 5%, 1%, 0.5% and 0.1%. Seeds from each sample were crushed, and then DNA extraction was performed by the CTAB 2% method. Using the immunochromatographic strip, it was possible to evaluate the expression of proteins related to the VT-Pro, Roundup Ready and Herculex events when whole seeds were used at the 0.2% level of contamination, whereas by the conventional PCR technique, it was possible to detect the TC1507 event in samples with 1% contamination.