Biologia

, Volume 73, Issue 1, pp 21–29 | Cite as

Development of a multiplex fluorescence quantitative PCR for detection of genetically modified organisms

  • Fengjun Wang
  • Junli Feng
  • Sudan Ye
  • Hannian Huang
  • Xianglin Zhang
Original Article

Abstract

The commercially available genetically modified plants authorized worldwide and therefore the target sequences for molecular detection of genetically modified organisms (GMOs) are ever-increasing. The European Union has implemented a set of very strict procedures for approval to grow, import and/or utilize GMOs as food or food ingredients. As a result, GMO laboratories and food production industry currently are forced to apply different methods to test raw material and complex processed food products. Three exogenous genes (the 35 s promoter of the cauliflower mosaic virus (35 s), nos terminator from Agrobacterium tumefaciens (nos), and the neomycin phosphotransferase II (nptII) gene) are commonly used in GMO detection. In this paper, a multiplex quantitative real-time PCR (qPCR) system was developed which allows simultaneously detection of the three exogenous genes in one reaction tube. The determined limits for the multiplex qPCR assays were 4 copies/reaction in maize samples. The specificity of the assays was demonstrated to be 100% according to the detection results of 23 genetically modified (GM) crops and 97 complex processed food products. The validation data show the individual PCR efficiency was accredited with negligible impacts between three detection channels in 7500 fluorescence quantitative PCR machine. These results indicate that this high-throughput multiplex qPCR method which combined with a reference gene is feasible for screening of GMOs, even for the processed food.

Keywords

Detection Genetically modified organisms Multiplex qPCR Screening 

Notes

Acknowledgments

This work was supported by the Grant from the Science Research Project of Zhejiang Federation of Supply and Marketing Cooperative under grant (No. 16SSY04); the Science Research Project of Zhejiang Institute of Economic and Trade under grant (No. 16QN02).

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Copyright information

© Section Botany: © Plant Science and Biodiversity Centre, Slovak Academy of Sciences 2018

Authors and Affiliations

  • Fengjun Wang
    • 1
  • Junli Feng
    • 2
  • Sudan Ye
    • 1
  • Hannian Huang
    • 1
  • Xianglin Zhang
    • 3
  1. 1.Department of Applied EngineeringZhejiang Institute of Economic and TradeHangzhouPeople’s Republic of China
  2. 2.Institute of SeafoodZhejiang Gongshang UniversityHangzhouPeople’s Republic of China
  3. 3.Xinjiang Entry-Exit Inspection and Quarantine BureauUrumqiPeople’s Republic of China

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