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2009 | 53 | 4 |

Tytuł artykułu

Validation of real-time PCR methods for the quantification of genetically-modified maize and soybean

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
The objective of this study was to validate TaqMan real-time PCR procedures for the quantification of genetically-modified maize (events: MON810, Bt11, Bt176, and T25) and soybean (event: GTS 40-3-2) with the capillary LightCycler 2.0 instrument. The quantification was done by the amplification of reference maize genes: hmg (MON810, Bt11), zSSIIb (Bt176), and adh1 (T25), and genetically-modified DNA fragments containing a part of the maize genome and modified genes at 5' end (MON810, Bt11, T25), or unique DNA sequence of genes (cryIAb/IVS9) used for the modification (Bt176). As regards GM soybean event GTS 40-3-2, soya specific lectin gene and GM construct-specific DNA fragment (CaMV35S/ CTP- EPSPS) were amplified in real-time PCR reactions. Real-time PCR performance criteria achieved in this validation study proved that it is possible to use the reaction protocol made for 96-well plates real-time PCR instruments at capillary Roche thermocycler. Result of the performance criteria obtained for real-time PCR validation study for the quantification of genetically-modified maize or soybean were satisfactory, close or ever better than the values of inter-laboratory ring-trials.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

53

Numer

4

Opis fizyczny

p.740-746,ref.

Twórcy

autor
  • Department of Hygiene of Animal Feedingstuffs, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Hygiene of Animal Feedingstuffs, National Veterinary Research Institute, 24-100 Pulawy, Poland

Bibliografia

  • 1. Alexander T.W., Reuter T., Aulrich K., Sharma R., Okine E.K., Dixon W.T., McAllister T.A.: A review of the detection and fate of novel plant molecules derived from biotechnology in livestock production. Anim Feed Sci Technol 2007, 133, 31-62.
  • 2. Anklam E., Gadani F., Heinze P., Pijnenburg H., Van den Eede G.: Analytical methods for detection and determination of genetically-modified organisms in agricultural crops and plant-derived food products. Eur Food Res Technol 2002, 214, 3-26.
  • 3. Deisingh A.K., Badrie N.: Detection approaches for genetically-modified organisms in foods. Food Res Int 2005, 38, 639-649.
  • 4. EN ISO 21570: 2007, Foodstuffs - Methods of analysis for the detection of genetically-modified organisms and derived products. Quantitative nucleic acid based methods.
  • 5. EN ISO 21571: 2005, Foodstuffs - Methods of analysis for the detection of genetically-modified organisms and derived products. Nucleic acid extraction.
  • 6. Higuchi R., Dollinger G., Walsh P. S., Griffith R.: Simultaneous amplification and detection of specific DNA sequences. Bio/Technol 1992, 10, 413-417.
  • 7. Hoist-Jensen A., Ronning S.B., Lovseth A.: PCR technology for screening and quantification of genetically-modified organisms (GMOs). Anal Bioanal Chem 2003, 375, 985-993.
  • 8. http://bgmo.jrc. ec.europa.eu/home/ict/methodsdatabase.h tm
  • 9. http://gmo-crl.jrc.ec.europa.eu
  • 10. Hubner P., Waiblinger U., Pietsch K., Brodmann P.: Validation of PCR methods for quantitation of genetically-modified plants in food. J AOAC Int 2001, 84, 1855-1864.
  • 11. Kuribara H., Shindo Y., Matsuoka T., Takubo K., Futo S., Aoki N., Hirao T., Akiyama H., Goda Y., Toyoda M., Hino A.: Novel reference molecules for quantitation of genetically-modified maize and soybean. J AOAC Int 2002, 85, 1077-1089.
  • 12. Kwiatek K., Sieradzki Z., Mazur M.: Prevalence of genetically-modified crops in animal feedingstuffs in Poland - Three year studies. Bull Vet Inst Pulawy 2007, 51, 609-613.
  • 13. Regulation (EC) No. 1830/2003 of the European Parliament and of the Council of 22 September 2003 concerning the traceability and labelling of genetically- modified organism and the traceability of the food and feed products produced from genetically-modified organism and amending Directive 2001/18/EC, OJ L 2003, 268, pp. 24-28.
  • 14. Regulation (EC) No. 1829/2003 of the European Parliament and of the Council of 22 September 2003 on genetically-modified food and feed, OJ L 2003, 268, pp. 1-23.
  • 15. Ronning S.B., Vaitilingom M., Berdal K.G., Holst- Jensen A.: Event specific quantitative PCR for genetically-modified Bt11 maize (Zea mays). Eur Food Res Technol 2003, 216, 347-354.
  • 16. Shindo Y., Kuribara H., Matsuoka T., Futo S., Sawada Ch., Shono J., Akiyama H., Goda Y., Toyoda M., Hino A.: Validation of PCR analyses for line-specific quantitation of genetically-modified maize and soybean using new reference molecules. J AOAC Int 2002, 85, 1119-1126.
  • 17. Sieradzki Z., Walczak M., Kwiatek K.: Occurrence of genetically-modified maize and soybean in animal feedingstuffs. Bull Vet Inst Pulawy 2006, 50, 359-362.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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