Display options
Cite
Datukishvili N, Kutateladze T, Gabriadze I, et al. New multiplex PCR methods for rapid screening of genetically modified organisms in foods. Front Microbiol. 2015;6:757doi: 10.3389/fmicb.2015.00757.
Datukishvili, N., Kutateladze, T., Gabriadze, I., Bitskinashvili, K., & Vishnepolsky, B. (2015). New multiplex PCR methods for rapid screening of genetically modified organisms in foods. Frontiers in microbiology, 6757. https://doi.org/10.3389/fmicb.2015.00757
Datukishvili, Nelly, et al. "New multiplex PCR methods for rapid screening of genetically modified organisms in foods." Frontiers in microbiology vol. 6 (2015): 757. doi: https://doi.org/10.3389/fmicb.2015.00757
Datukishvili N, Kutateladze T, Gabriadze I, Bitskinashvili K, Vishnepolsky B. New multiplex PCR methods for rapid screening of genetically modified organisms in foods. Front Microbiol. 2015 Jul 24;6:757. doi: 10.3389/fmicb.2015.00757. eCollection 2015. PMID: 26257724; PMCID: PMC4513241.
Copy Download .nbib Format: NLM
Share it on

Front Microbiol. 2015 Jul 24;6:757. doi: 10.3389/fmicb.2015.00757. eCollection 2015.

New multiplex PCR methods for rapid screening of genetically modified organisms in foods.

Frontiers in microbiology

Nelly Datukishvili, Tamara Kutateladze, Inga Gabriadze, Kakha Bitskinashvili, Boris Vishnepolsky

Affiliations

  1. Ivane Beritashvili Center of Experimental Biomedicine Tbilisi, Georgia ; Faculty of Natural Sciences and Engineering, Ilia State University Tbilisi, Georgia.
  2. Ivane Beritashvili Center of Experimental Biomedicine Tbilisi, Georgia.
  3. Faculty of Natural Sciences and Engineering, Ilia State University Tbilisi, Georgia.

PMID: 26257724 PMCID: PMC4513241 DOI: 10.3389/fmicb.2015.00757

Abstract

We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs). New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV) 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS) terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps) gene and 258 bp fragment of Cry1Ab delta-endotoxin (cry1Ab) gene for GMO screening. The certified reference materials containing Roundup Ready soybean (RRS) and maize MON 810 were applied for the development and optimization of uniplex and multiplex PCR systems. Evaluation of amplification products by agarose gel electrophoresis using negative and positive controls confirmed high specificity and sensitivity at 0.1% GMO for both RRS and MON 810. The fourplex PCR was developed and optimized that allows simultaneous detection of three common transgenic elements, such as: CaMV 35S promoter, NOS terminator, epsps gene together with soybean-specific lectin gene. The triplex PCR developed enables simultaneous identification of transgenic elements, such as: 35S promoter and cry1Ab gene together with maize zein gene. The analysis of different processed foods demonstrated that multiplex PCR methods developed in this study are useful for accurate and fast screening of GM food products.

Keywords: DNA marker; food; genetically modified organism; multiplex PCR; screening

References

  1. J AOAC Int. 2009 May-Jun;92(3):765-72 - PubMed
  2. Biotechnol Adv. 2009 Nov-Dec;27(6):1071-82 - PubMed
  3. BMC Bioinformatics. 2012 Jun 18;13:134 - PubMed
  4. Anal Bioanal Chem. 2010 Mar;396(6):1991-2002 - PubMed
  5. J Agric Food Chem. 2003 Sep 24;51(20):5829-34 - PubMed
  6. Anal Bioanal Chem. 2010 Mar;396(6):2113-23 - PubMed
  7. Shokuhin Eiseigaku Zasshi. 2001 Feb;42(1):24-32 - PubMed
  8. Biotechnol Adv. 2012 Nov-Dec;30(6):1318-35 - PubMed
  9. Bioinformatics. 2005 Apr 15;21(8):1701-2 - PubMed
  10. Anal Biochem. 2003 Dec 15;323(2):164-70 - PubMed
  11. Comput Biol Chem. 2009 Jun;33(3):235-8 - PubMed
  12. Plant Mol Biol. 2006 May;61(1-2):123-39 - PubMed
  13. J Biomed Biotechnol. 2012;2012:402418 - PubMed
  14. J Agric Food Chem. 2008 Dec 24;56(24):11596-606 - PubMed
  15. J Integr Plant Biol. 2011 Jul;53(7):539-51 - PubMed
  16. Anal Bioanal Chem. 2014 Oct;406(26):6485-97 - PubMed
  17. Sci Rep. 2015 Jan 05;5:7616 - PubMed
  18. Anal Bioanal Chem. 2008 Oct;392(3):369-84 - PubMed
  19. Anal Bioanal Chem. 2008 Oct;392(3):355-67 - PubMed
  20. J Agric Food Chem. 2005 Dec 14;53(25):9713-21 - PubMed
  21. Genomics. 2011 Aug;98(2):137-44 - PubMed
  22. BMC Bioinformatics. 2008 Jun 04;9:260 - PubMed
  23. J Agric Food Chem. 2004 Jun 2;52(11):3275-80 - PubMed

Publication Types