Analytical and Bioanalytical Chemistry

, Volume 398, Issue 5, pp 2279–2288 | Cite as

A novel, sensitive and specific real-time PCR for the detection of traces of allergenic Brazil nut (Bertholletia excelsa) in processed foods

Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

Food-allergic individuals have to strictly avoid the offending food because no causative immunotherapies are available. Thus, reliable labelling of allergenic ingredients or precautionary labelling of cross-contacts with potential allergens is of major importance. Verification of compliance with labelling requirements and identification of cross-contacts demand test methods that enable the specific and sensitive detection of the analyte. Brazil nut (Bertholletia excelsa) is such a food commodity with allergenic potential. We describe the development of a novel qualitative real-time polymerase chain reaction (PCR) specific for Brazil nut DNA and its comparison with a qualitative commercially available lateral flow device (LFD) that detects Brazil nut protein. Specificity was investigated with 58 foods, and no false-positive reactions were observed in real-time PCR. The sensitivity was investigated with spiked chocolate and incurred dough samples as well as cookies baked thereof. The simultaneous spiking of matrices with identical amounts of Brazil nut and peanut between 5 and 100,000 mg/kg allowed the verification of the spike quality with two peanut-specific enzyme-linked immunosorbent assay. The real-time PCR detected Brazil nut in all three matrices down to the lowest investigated spike level of 5 mg/kg. The real-time PCR results from the analysis of 15 retail samples were confirmed by LFD results and were in concordance with the labelling of products. The real-time PCR showed unparalleled specificity, and primary data indicated potentially quantitative features in spiked and retail samples. Because of entirely reproducible chemistry of this real-time PCR, this is the first generally available Brazil nut-specific detection method with an appropriate sensitivity to help avoid severe allergic reactions for Brazil nut-allergic individuals.

Keywords

Brazil nut Bertholletia excelsa Allergen labelling ELISA Real-time PCR Lateral flow device (LFD) 
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Notes

Acknowledgement

Leaves for DNA extraction were kindly provided by the botanical garden of the Johann Wolfgang Goethe University Frankfurt and the Palmengarten Frankfurt. Kornelia Kleiner is acknowledged for her help in sample spiking.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Martin Röder
    • 1
  • Helene Filbert
    • 1
  • Thomas Holzhauser
    • 1
  1. 1.Division of AllergologyPaul-Ehrlich-InstitutLangenGermany

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