A novel quadruplex real-time PCR method for simultaneous detection of Cry2Ae and two genetically modified cotton events (GHB119 and T304-40).

Xiang Li,Yueming Liu,Liangwen Pan,Jielin Yang,Yuping He,Xiuxiu Wang

doi:10.1186/1472-6750-14-43

Abstract

BackgroundTo date, over 150 genetically modified (GM) crops are widely cultivated. To comply with regulations developed for genetically modified organisms (GMOs), including labeling policies, many detection methods for GMO identification and quantification have been developed.ResultsTo detect the entrance and exit of unauthorized GM crop events in China, we developed a novel quadruplex real-time PCR method for simultaneous detection and quantification of GM cotton events GHB119 and T304-40 in cotton-derived products (based on the 5′-flanking sequence) and the insect-resistance gene Cry2Ae. The limit of detection was 10 copies for GHB119 and Cry2Ae and 25 copies for T304-40. The limit of quantification was 25 copies for GHB119 and Cry2Ae and 50 copies for T304-40. Moreover, low bias and acceptable standard deviation and relative standard deviation values were obtained in quantification analysis of six blind samples containing different GHB119 and T304-40 ingredients.ConclusionsThe developed quadruplex quantitative method could be used for quantitative detection of two GM cotton events (GHB119 and T304-40) and Cry2Ae gene ingredient in cotton derived products.

Highlights

IntroductionOver 150 genetically modified (GM) crops are widely cultivated. To comply with regulations developed for genetically modified organisms (GMOs), including labeling policies, many detection methods for GM organisms (GMO) identification and quantification have been developed
To date, over 150 genetically modified (GM) crops are widely cultivated
The developed quadruplex quantitative method could be used for quantitative detection of two GM cotton events (GHB119 and T304-40) and Cry2Ae gene ingredient in cotton derived products

Summary

Introduction

Over 150 genetically modified (GM) crops are widely cultivated. To comply with regulations developed for genetically modified organisms (GMOs), including labeling policies, many detection methods for GMO identification and quantification have been developed. As the quantity of exogenous genes and GM crop events increase, several multiplex methods for simultaneous detection of more than one genetic element have been developed, such as multiplex PCR, microarray analysis, suspension array analysis, microdroplet-based PCR and others [4,5,6]. These techniques are especially useful for detecting GM events in complex or mixed samples that contain multiple exogenous insertions. Only a few real-time multiplex PCR methods are reported and maximum four targets are simultaneously detected [10,11]

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Results

Conclusion