A real-time immuno-PCR assay for the detection of transgenic Cry1Ab protein

The adaptation and production of transgenic crops harboring Cry1Ab protein is increasing every year globally due to their potent insecticidal activity. The Cry1Ab protein produced by Bacillus thuringiensis confers resistance against several lepidopteran insect pests. The release of transgenic crops/produce in the market worldwide has increased the need of regulatory affairs to monitor and verify the presence of transgenic protein in crops/produce. In this regard, the real-time immuno-PCR (IPCR) assay was developed for the detection and quantification of Cry1Ab protein. The IPCR assay showed high sensitivity with minimum detection limit of 100 pg/mL (0.1 ppb) and found to be 10 times more sensitive than sandwich ELISA. Under the optimized assay conditions, Cry1Ab protein can be determined in the concentration ranged from 100 pg/mL to 100 ng/mL. As results suggest, this assay could be a powerful tool for the detection of even trace amounts of Cry1Ab protein in transgenic crops.