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

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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.

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HMW glutenin:

High molecular weight glutenin


Polymerase chain reaction




Yakima Yellow






IUB-code for cytosin, guanin or thymin


IUB-code for cytosin or thymin


  1. Taylor SL (1987) Allergic and sensitivity reactions to food components. In: Hathcock JN (ed) Nutritional toxicology, vol II. Academic, New York, pp 173–197

    Google Scholar 

  2. Holgate ST, Church MK, Lichtenstein LM (2001) Allergy, 2nd edn edn. Mosby, St Louis

    Google Scholar 

  3. Deutsche Zöliakiegesellschaft (2007)

  4. Schultz M, Hellerbrand C (2002) Dtsch Arzteblt 99(28–29):A1949

    Google Scholar 

  5. Janatuinen EK, Pikkarainen PH, Kemppainen TA, Kosma VM, Järvinen RMK, Uusitupa MIJ, Julkunen RJK (1995) N Engl J Med 333:1033–1037

    Article  CAS  Google Scholar 

  6. Janatuinen EK, Kemppainen TA, Pikkarainen PH, Holm KH, Kosma VM, Uusitupa MIJ, Mäki M, Julkunen RJK (2000) Gut 46:327–331

    Article  CAS  Google Scholar 

  7. Janatuinen EK, Kemppainen TA, Julkunen RJK, Kosma VM, Mäki M, Heikkinen M, Uusitupa MIJ (2002) Gut 50:332–335

    Article  CAS  Google Scholar 

  8. Anonymous (2002) Deutsche Forschungsanstalt für Lebensmittelchemie; Jahresbericht 2002. Available from http: //

  9. Kilmartin C, Lynch S, Abuzakouk M, Wieser H, Feighery C (2003) Gut 52:47–52

    Article  CAS  Google Scholar 

  10. Pasternack A (2006) Labor and more volume 1/2006:18

  11. Anonymus Codex Alimentarius: CODEX STAN 118. ALINORM 08/31/26, Appendix III

  12. Pöpping B, Holzhauser T (2004) Dtsch Lebensmitt Rundsch 8:285–293

    Google Scholar 

  13. Valdés I, García E, Llorente M, Méndez E (2003) Eur J Gastroenterol Hepatol 15:465–474

    Article  Google Scholar 

  14. Méndez E, Vela C, Immer U, Janssen FW (2005) Eur J Gastroenterol Hepatol 17:1053–1063

    Article  Google Scholar 

  15. Terzia V, Malnati M, Barbanera M, Stanca AM, Faccioli P (2003) J Cereal Sci 38:87–94

    Article  Google Scholar 

  16. Dahinden I, von Buren M, Lüthy J (2001) Eur Food Res Technol 212:228–233

    Article  CAS  Google Scholar 

  17. Sandberg M, Lundberg L, Ferm M, Malmheden Yman I (2003) Eur Food Res Technol 217:344–349

    Article  CAS  Google Scholar 

  18. Anonymous (2005) International Standard EN ISO 22174

  19. Rogers S, Bendich A (1985) Plant Mol Biol 5:69–76

    Article  CAS  Google Scholar 

  20. Anonymous (2005) International Standard EN ISO 21571

  21. Reichardt M, Rogers S (1994) In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidmann JG, Smith JA, Struhl K (eds), supplement 27. Wiley, New York. doi 10.1002/mrd.1080010210

  22. Benson DA, Karsch-Mizrachi I, Lipman DJ, Ostell J, Wheeler DL (2007) Nucleic Acids Res 35:D21–D25

    Article  CAS  Google Scholar 

  23. Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG, Thompson JD (2003) Nucleic Acids Res 31:3497–3500

    Article  CAS  Google Scholar 

  24. Hernandez M, Esteve T, Pla M (2005) J Agric Food Chem 53:7003–7009

    Article  CAS  Google Scholar 

  25. Allmann M, Candrian U, Ch Höfelein, Lüthy J (1993) Z Lebensm Unters Forsch 196:248–251

    Article  CAS  Google Scholar 

  26. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DL (1997) Nucleic Acids Res 25:3389–3402

    Article  CAS  Google Scholar 

  27. Meyer R, Candrian U, Lüthi J (1993) Mitt Geb Lebensmittelunters Hyg 84:112–121

    CAS  Google Scholar 

  28. Rossen L, Norskov P, Holmstrom K, Rasmussen OF (1992) Int J Food Microbiol 17:37–45

    Article  CAS  Google Scholar 

  29. Meyer R, Höfelein C, Lüthi J, Candrian U (1995) J AOAC Int 78:1542–1557

    CAS  Google Scholar 

  30. Hupfer C, Hotzel H, Sachse K, Engel K-H (1998) Z Lebensm Unters Forsch A 205:442–445

    Article  Google Scholar 

  31. Poser R, Detsch R, Fischer K, Müller WD, Behrschmidt M, Schwägele F (2000) Fleischwirtschaft 8:87–89

    Google Scholar 

  32. Anonymous (2007) prEN 15634-1:2007. Norm Entwurf. Deutsches Institut für Normung, Berlin

  33. Rooney AP, Ward TJ (2005) PNAS 102:5084–5089

    Article  CAS  Google Scholar 

  34. Zoschke R, Liere K, Börner T (2007) Plant J 50:710–722

    Article  CAS  Google Scholar 

  35. Anklam E, Gadani F, Heinze P, Pijnenburg H, van den Eede G (2002) Eur Food Res Technol 214:3–16

    Article  CAS  Google Scholar 

  36. Moreano Guerra FX (2005) Thesis. Technische Universität, München. URL:

  37. Forde J, Malpica JM, Halford NG, Shewry PR, Anderson OD, Greene FC, Miflin BJ (1985) Nucleic Acids Res 13:6817–6832

    Article  CAS  Google Scholar 

  38. Bennett MD, Smith JB (1976) Philos Trans R Soc Lond B Biol Sci 274:227–274

    Article  CAS  Google Scholar 

  39. Arumuganathan K, Earle ED (1991) Plant Mol Biol Rep 9:211–215

    Google Scholar 

  40. Pauli U, Liniger M, Zimmermann A, Schrott M (2000) Mitt Lebensmitteluntersuchung Hyg 91:491–501

    CAS  Google Scholar 

  41. Tilley M (2004) Cereal Chem 81:44–47

    Article  CAS  Google Scholar 

  42. Anonymous (2007) Codex Alimentarius—Codex Committee on Nutrition and Foods for Special Dietary CX/NFSDU 07/29/4 Add 1, Bonn, Germany

  43. Belitz HD, Grosch W, Schieberle A (2001) Getreideprodukte in Lehrbuch der Lebensmittelchemie, 5 edn edn. Springer, Berlin, p 656

    Google Scholar 

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Correspondence to Dietrich Maede.

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Zeltner, D., Glomb, M.A. & Maede, D. Real-time PCR systems for the detection of the gluten-containing cereals wheat, spelt, kamut, rye, barley and oat. Eur Food Res Technol 228, 321–330 (2009).

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