Analytical and Bioanalytical Chemistry

, Volume 407, Issue 16, pp 4829–4834 | Cite as

GMO detection in food and feed through screening by visual loop-mediated isothermal amplification assays

  • Cong Wang
  • Rong Li
  • Sheng Quan
  • Ping Shen
  • Dabing Zhang
  • Jianxin Shi
  • Litao Yang


Isothermal DNA/RNA amplification techniques are the primary methodology for developing on-spot rapid nucleic acid amplification assays, and the loop-mediated isothermal amplification (LAMP) technique has been developed and applied in the detection of foodborne pathogens, plant/animal viruses, and genetically modified (GM) food/feed contents. In this study, one set of LAMP assays targeting on eight frequently used universal elements, marker genes, and exogenous target genes, such as CaMV35S promoter, FMV35S promoter, NOS, bar, cry1Ac, CP4 epsps, pat, and NptII, were developed for visual screening of GM contents in plant-derived food samples with high efficiency and accuracy. For these eight LAMP assays, their specificity was evaluated by testing commercial GM plant events and their limits of detection were also determined, which are 10 haploid genome equivalents (HGE) for FMV35S promoter, cry1Ac, and pat assays, as well as five HGE for CaMV35S promoter, bar, NOS terminator, CP4 epsps, and NptII assays. The screening applicability of these LAMP assays was further validated successfully using practical canola, soybean, and maize samples. The results suggested that the established visual LAMP assays are applicable and cost-effective for GM screening in plant-derived food samples.


Genetically modified organism Loop-mediated isothermal amplification Screening test 

Supplementary material

216_2015_8652_MOESM1_ESM.pdf (977 kb)
ESM 1(PDF 976 kb)


  1. 1.
    James C (2014) Global status of commercialized biotech/GM crops: 2014. ISAAA Brief 49Google Scholar
  2. 2.
    Miraglia M, Berdal KG, Brera C, Corbisier P, Holst-Jensen A, Kok EJ, Marvin HJ, Schimmel H, Rentsch J, van Rie JP, Zagon J (2004) Detection and traceability of genetically modified organisms in the food production chain. Food Chem Toxicol 42:1157–80CrossRefGoogle Scholar
  3. 3.
    Broeders SR, De Keersmaecker SC, Roosens NH (2012) How to deal with the upcoming challenges in GMO detection in food and feed. J Biomed Biotechnol 2012:11Google Scholar
  4. 4.
    (2003) European Parliament; Council of the European Union. Commission regulation (EC) No 1830/2003. Concerning the traceability and labeling of genetically modified organisms and the traceability of food and feed products produced from genetically modified organisms and amending Directive 2001/18/EC. Off J Eur Union 268:24-28Google Scholar
  5. 5.
    Bean CE (2002) Japan Biotechnology MAFF’s Biotech food labeling standards (revised). GAIN Report JA2010, USDA, GAINGoogle Scholar
  6. 6.
    Ministry of Agriculture and Forestry of Korea (2000) Guidelines for labeling of genetically modified agricultural products. MAF Notification No. 2000-31Google Scholar
  7. 7.
    Walker G, Fraiser M, Schram J, Little MC (1992) Strand displacement amplification-an isothermal, in vitro DNA amplification technique. Nucleic Acids Res 20:1691–1696CrossRefGoogle Scholar
  8. 8.
    Cao Y, Kim HJ, Li Y, Kong H, Lemieux B (2013) Helicase-dependent amplification of nucleic acids. Curr Protoc Mol Biol 104:Unit 15.11Google Scholar
  9. 9.
    Lizardi PM, Huang X, Zhu Z, Bray-Ward P, Thomas DC, Ward DC (1998) Mutation detection and single-molecule counting using isothermal rolling-circle amplification. Nat Genet 19:225–32CrossRefGoogle Scholar
  10. 10.
    Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:e63CrossRefGoogle Scholar
  11. 11.
    Lee D, La Mura M, Allnutt TR, Powell W (2009) Detection of genetically modified organisms (GMOs) using isothermal amplification of target sequences. BMC Biotechnol 9:7CrossRefGoogle Scholar
  12. 12.
    Guan X, Guo J, Shen P, Yang L, Zhang D (2010) Visual and rapid detection of two genetically modified soybean events using loop-mediated isothermal amplification method. Food Anal Methods 3:313–320CrossRefGoogle Scholar
  13. 13.
    Holst-Jensen A, Rønning S, Løvseth A, Berdal K (2003) PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal Bioanal Chem 375:985–993Google Scholar
  14. 14.
    Chen L, Guo J, Wang Q, Kai G, Yang L (2011) Development of the visual loop-mediated isothermal amplification assays for seven genetically modified maize events and their application in practical samples analysis. J Agric Food Chem 59:5914–5918CrossRefGoogle Scholar
  15. 15.
    Kiddle G, Hardinge P, Buttigieg N (2012) GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use. BMC Biotechnol 12:15CrossRefGoogle Scholar
  16. 16.
    Guo J, Yang L, Chen L, Morisset D, Li X, Pan L, Zhang D (2011) MPIC: a high-throughput analytical method for multiple DNA targets. Anal Chem 83:1579–1586CrossRefGoogle Scholar
  17. 17.
    Shao N, Jiang S, Zhang M, Wang J, Guo S, Li Y, Jiang H, Liu C, Zhang D, Yang L, Tao S (2014) MACRO: a combined microchip-PCR and microarray system for high-throughput monitoring of genetically modified organisms. Anal Chem 86:1269–1276CrossRefGoogle Scholar
  18. 18.
    Zhang M, Liu Y, Chen L, Quan S, Jiang S, Zhang DB, Yang LT (2013) One simple dna extraction device and its combination with modified visual loop-mediated isothermal amplification for rapid on-field detection of genetically modified organisms. Anal Chem 85:75–82CrossRefGoogle Scholar
  19. 19.
    Yang L, Pan A, Zhang K, Guo J, Yin C, Chen J, Huang C, Zhang D (2005) Identification and quantification of three genetically modified insect resistant cotton lines conventional and taqman real-time polymerase chain reaction methods. J Agric Food Chem 53:6222–6229CrossRefGoogle Scholar
  20. 20.
    Broeders S, Huber I, Grohmann L, Berben G, Taverniers I, Mazzara M, Roosens N, Morisset D (2014) Guidelines for validation of qualitative real-time PCR methods. Trends Food Sci Technol 37:115–126CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Cong Wang
    • 1
  • Rong Li
    • 1
  • Sheng Quan
    • 1
  • Ping Shen
    • 2
  • Dabing Zhang
    • 1
  • Jianxin Shi
    • 1
  • Litao Yang
    • 1
  1. 1.Collaborative Innovation Center for Biosafety of GMOs, National Center for the Molecular Characterization of Genetically Modified Organisms, School of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Development Center of Science and TechnologyMinistry of Agriculture of People’s Republic of ChinaBeijingChina

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