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European Food Research and Technology

, Volume 228, Issue 3, pp 321–330 | Cite as

Real-time PCR systems for the detection of the gluten-containing cereals wheat, spelt, kamut, rye, barley and oat

  • Daniela Zeltner
  • Marcus A. Glomb
  • Dietrich MaedeEmail author
Original Paper

Abstract

Real-time PCR assays, using TaqMan® probes, were applied to detect the gluten-containing cereals. Homologues target sequences encoding high molecular weight (HMW) glutenin were chosen to detect wheat, kamut, spelt and rye. For detecting barley, the gene Hor3 was selected. For the detection of oat the gene encoding the 12S seed storage protein was chosen. Based on this sequence data, primer and probe sequences were generated for the real-time PCR. A plant specific primer probe system based on 18S rRNA gene was chosen to detect amplificability of the extracted nucleic acids. The specificity of the primer and probe systems was checked using different lines from different origins of the species to be detected. The HMW glutenin system is specific for the corresponding species, as are the systems for the barley Hor3 gene and the oat 12S seed storage protein. The sensitivity of the systems was determined testing different matrices. With the HMW glutenin system 2.5 mg/kg of wheat in vegetable food matrices and 5 mg/kg of wheat in meat products were detected. The oat and the barley specific systems resulted in a sensitivity of 10 mg/kg. The detection method showed a satisfactory ruggedness.

Keywords

Molecular detection Real-time PCR Gluten-containing cereals Wheat Rye Barley Oat 

Abbreviations

HMW glutenin

High molecular weight glutenin

PCR

Polymerase chain reaction

FAM

6-Carboxyfluorescein

YAK

Yakima Yellow

TMR (TAMRA)

6-Carboxy-tetramethylrhodamine

DB

Dabcyl

B

IUB-code for cytosin, guanin or thymin

Y

IUB-code for cytosin or thymin

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Daniela Zeltner
    • 1
  • Marcus A. Glomb
    • 2
  • Dietrich Maede
    • 1
    Email author
  1. 1.LAV Sachsen-AnhaltHalle (Saale)Germany
  2. 2.Martin-Luther-Universität Halle-WittenbergHalle (Saale)Germany

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