Skip to main content
SpringerLink
Log in

Detection system of stacked genetically modified maize using multiplex PCR

  • Research Article
  • Published:
Food Science and Biotechnology Aims and scope Submit manuscript

Abstract

A multiplex polymerase chain reaction (PCR) method was developed to identify and distinguish 3 kinds of stacked genetically modified (GM) maize (MON810× MON863, NK603×MON863, and NK603×MON810× MON863). Four primer pairs, SSIIb JHF/JHR, C3b 5′/TAP1–3′, HS01/cry-CR01, and HS01/CTP164-3′ yielded 101, 129, 194, and 314 bp amplicons, respectively, Using the genomic DNA of the 3 stacked GM maize as templates, 3 or 4 corresponding PCR amplicons were amplified with similar band intensities by the multiplex PCR. The limit of detection (LOD) was approximately 0.5% for 3 kinds of stacked GM maize, using the multiplex PCR. The detection system using multiplex PCR developed in this study may be applicable to monitoring, identifying, and distinguishing not only the stacked GM maizes but also other stacked genetically modified organisms (GMOs).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Elenis DS, Kalogianni DP, Glynou K, Ioannou PC, Christopoulos TK. Advances in molecular techniques for the detection and quantification of genetically modified organisms. Anal. Bioanal. Chem. 392: 347–354 (2008)

    Article  CAS  Google Scholar 

  2. Engel KH, Frenzel T, Miller A. Current and future benefits from the use of GM technology in food production. Toxicol. Lett. 127: 329–336 (2002)

    Article  CAS  Google Scholar 

  3. Chandler SF, Lu C-Y. Biotechnology in ornamental horticulture. In Vitro Cell. Dev.-Pl 41: 591–601 (2005)

    Article  Google Scholar 

  4. Stephenson JR, Warnes A. Release of genetically modified microorganisms into the environment. J. Chem. Technol. Biot. 65: 5–14 (1996)

    Article  CAS  Google Scholar 

  5. Kim HY, Kim JH, Oh MH. Regulation and detection methods for genetically modified foods in Korea. Pure Appl. Chem. 82: 129–137 (2010)

    Article  CAS  Google Scholar 

  6. Michelini E, Simoni P, Cevenini L, Mezzanotte L, Roda A. New trends in bioanalytical tools for the detection of genetically modified organisms: An update. Anal. Bioanal. Chem. 392: 355–367 (2008)

    Article  CAS  Google Scholar 

  7. Yang L, Pan A, Jia J, Ding J, Chen J, Cheng H, Zhang C, Zhang D. Validation of a tomato-specific gene, LAT52, used as an endogenous reference gene in qualitative and real-time quantitative PCR detection of transgenic tomatoes. J. Agr. Food Chem. 53: 183–190 (2005)

    Article  CAS  Google Scholar 

  8. Forte VT, Di Pinto A, Martino C, Tantillo GM, Grasso G, Schena FP. A general multiplex-PCR assay for the general detection of genetically modified soya and maize. Food Control 16: 535–539 (2005)

    Article  CAS  Google Scholar 

  9. Vollenhofer S, Burg K, Schmidt J, Kroath H. Genetically modified organisms in food-screening and specific detection by polymerase chain reaction. J. Agr. Food Chem. 47: 5038–5043 (1999)

    Article  CAS  Google Scholar 

  10. Allmann M, Candrian U, Höfelein C, Lüthy J. Polymerase chain reaction (PCR): A possible alternative to immunochemical methods assuring safety and quality of food: Detection of wheat contamination in non-wheat food products. Z. Lebensm. Unters. For. 196: 248–251 (1993)

    Article  CAS  Google Scholar 

  11. Meyer R, Chardonnens F, Hübner P, Lüthy J. Polymerase chain reaction (PCR) in the quality and safety assurance of food: Detection of soya in processed meat products. Z. Lebensm. Unters. For. 203: 339–344 (1996)

    Article  CAS  Google Scholar 

  12. Holst-Jensen A, Rønning SB, Løvseth A, Berdal KG. PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal. Bioanal. Chem. 375: 985–993 (2003)

    CAS  Google Scholar 

  13. De Schrijver A, Devos Y, Van den Bulcke M, Cadot P, De Loose M, Reheul D, Sneyers M. Risk assessment of GM stacked events obtained from crosses between GM events. Trends Food Sci. Tech. 18: 101–109 (2007)

    Article  Google Scholar 

  14. Akiyama H, Watanabe T, Wakabayashi K, Nakade S, Yasui S, Sakata K, Chiba R, Spiegelhalter F, Hino A, Maitani T. Quantitative detection system for maize sample containing combined-trait genetically modified maize. Anal. Chem. 77: 7421–7428 (2005)

    Article  CAS  Google Scholar 

  15. Lee B, Kim CG, Park JY, Park KW, Kim HJ, Yi H, Jeong SC, Yoon WK, Kim HM. Monitoring the occurrence of genetically modified soybean and maize in cultivated fields and along the transportation routes of the Incheon Port in South Korea. Food Control 20: 250–254 (2009)

    Article  Google Scholar 

  16. Xu WT, Yuan YF, Luo YB, Bai WB, Zhang CJ, Huang KL. Eventspecific detection of stacked genetically modified maize Bt11×GA21 by UP-M-PCR and real-time PCR. J. Agr. Food Chem. 57: 395–402 (2009)

    Article  CAS  Google Scholar 

  17. Rudi K, Rud I, Holck A. A novel multiplex quantitative DNA array based PCR (MQDA-PCR) for quantification of transgenic maize in food and feed. Nucleic Acids Res. 31: e62 (2003)

    Article  Google Scholar 

  18. Höhne M, Santisi CR, Meyer R. Real-time multiplex PCR: An accurate method for the detection and quantification of 35S-CaMV promoter in genetically modified maize-containing food. Eur. Food Res. Technol. 215: 59–64 (2002)

    Article  Google Scholar 

  19. Mano J, Shigemitsu N, Futo S, Akiyama H, Teshima R, Hino A, Furui S, Kitta K. Real-time PCR array as a universal platform for the detection of genetically modified crops and its application in identifying unapproved genetically modified crops in Japan. J. Agr. Food Chem. 57: 26–37 (2009)

    Article  CAS  Google Scholar 

  20. Xu J, Zhu S, Miao H, Huang W, Qiu M, Huang Y, Fu X, Li Y. Event-specific detection of seven genetically modified soybean and maizes using multiplex-PCR coupled with oligonucleotide microarray. J. Agr. Food Chem. 55: 5575–5579 (2007)

    Article  CAS  Google Scholar 

  21. Gruère GP, Carter CA, Hossein Farzin Y. Explaining international differences in genetically modified food labeling policies. Rev. Int. Econ. 17: 393–408 (2009)

    Article  Google Scholar 

  22. Kim JH, Kim HY. Event-specific detection methods for genetically modified maize MIR604 using real-time PCR. Food Sci. Biotechnol. 18: 1118–1123 (2009)

    CAS  Google Scholar 

  23. Kim JH, Park SH, Kim HY. Multiplex PCR detection of four events of GM maize (Event 3272, LY038, MIR162, and MON88017). J. Korean Soc. Appl. Biol. Chem. 52: 105–107 (2009)

    Article  CAS  Google Scholar 

  24. Matsuoka T, Kawashima Y, Akiyama H, Miura H, Goda Y, Kusakabe Y, Isshiki K, Toyoda M, Hino A. A method of detecting recombinant DNAs from four lines of genetically modified maize. J Food Hyg. Soc. Jpn. 41: 137–143 (2000)

    Article  CAS  Google Scholar 

  25. Kim JH, Song HS, Heo MS, Lee WY, Lee SH, Park SH, Park HK, Kim MC, Kim HY. Detection of eight different events of genetically modified maize by multiplex PCR method. Food Sci. Biotechnol. 15: 148–151 (2006)

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Life Sciences & Resources and Graduate School of Biotechnology, Kyung Hee University, Yongin, Gyeonggi, 446-701, Korea

    Su-Youn Kim, Jae-Hwan Kim, Hyungjae Lee & Hae-Yeong Kim

Authors
  1. Su-Youn Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jae-Hwan Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hyungjae Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hae-Yeong Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hae-Yeong Kim.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, SY., Kim, JH., Lee, H. et al. Detection system of stacked genetically modified maize using multiplex PCR. Food Sci Biotechnol 19, 1029–1033 (2010). https://doi.org/10.1007/s10068-010-0144-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10068-010-0144-5

Keywords

Search

Navigation