Quantitative Detection of Pest-resistant Genetically Modified Maize

doi:10.11924/j.issn.1000-6850.2013-1193

Abstract

Abstract: The aim was to build efficient, rapid and accurate event-specific quantitative real-time PCR detection methods of insect-resistant genetically modified maize MIR162. SYBR Green I and TaqMan Probe quantitative PCR methods were established in this study. The melting curves of MIR162 event-specific and endogenous reference gene zSSIIb were both single peak which mean the primers were specific. The recovery rates of SYBR Green I and TaqMan were between 90%-100% and their R2 values were both above 0.998. The limits of detection (LOD) and limits of quantitation (LOQ) of these two methods were 24 and 48 copies, respectively, which suggested that the two quantitative systems had an acceptable and high sensitivity. Moreover, the standard deviations (SD) and relative standard deviations (RSD) of repeatability ranged from 0.05 to 0.13 and 0.17% to 0.40% for SYBR Green I; 0.03 to 0.14 and 0.11% to 0.44% for TaqMan. Thereafter, three mixed maize samples (3%, 1%, 0.5%) were quantified to be 3.13%, 1.09%, 0.53% (SYBR Green I) and 3.07%, 1.02%, 0.50% (Taqman) respectively. T-test showed the quantification results of these two methods were not significantly different. The two event-specific quantitative PCR systems established in the studies were acceptable and suitable for MIR162 identification and quantification, and the users might select corresponding method based on actual demand and laboratory conditions.

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