Polymerase chain reaction coupled with peptide nucleic acid high-performance liquid chromatography for the sensitive detection of traces of potentially allergenic hazelnut in foodstuffs
作者： Andrea Germini , Elena Scaravelli , Francesca Lesignoli , Stefano Sforza , Roberto Corradini
DOI: 10.1007/S00217-004-1105-0
关键词:
摘要: A new DNA extraction method suitable for a wide variety of complex food matrices has been devised and applied in combination with polymerase chain reaction (PCR) the sensitive detection specific sequence univocally identifying presence potentially allergenic hazelnut (Corylus avellana). 156 base pair amplicon corresponding to an internal region complementary major allergen (Cor 1) was designed, found be highly specific; tested on both pure able confirm down 5 pg its DNA. The further confirmed by high-performance liquid chromatography (HPLC) analysis peptide nucleic acid (PNA) probe. 15-mer PNA probe expressly designed synthesized hybridize previously described amplicon. Given reported specificity high hybridization efficiency, used develop anion-exchange HPLC allowing fast reliable confirmation identity amplified products. PCR–HPLC successfully commercial samples, hidden allergens even products where as ingredient or possible contaminant not reported.
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