A procedure for olive oil traceability and authenticity: DNA extraction, multiplex PCR and LDR–universal array analysis
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Traceability of olive oils is relevant not only in assessing their origin, but also in protecting against frauds. Here, we present an improvement of the assay previously developed for the genotyping of forty-nine frequently grown Mediterranean olive cultivars by ligation detection reaction (LDR)/universal array (UA), refining the entire procedure in order to address DNA extracted from monovarietal olive oils. Firstly, a simple and robust protocol to extract amplifiable DNA from olive oil was developed. Then, the SNP-containing DNA sequences were simultaneous amplified by multiplex PCR and used on a LDR-UA platform, which gave precise and accurate genotype results. Thirteen out of the seventeen investigated SNPs were amplifiable in multiplex PCR, and were sufficient to unequivocally discriminate the forty-nine cultivars. The availability of this semi-automated SNP genotyping assay should help food testing laboratories to verify the origin and authenticity of monovarietal extra-virgin olive oils.
KeywordsOlive Monovarietal oil DNA extraction Single nucleotide polymorphisms (SNPs) Multiplex PCR (mPCR) Ligation detection reaction (LDR) Universal array (UA)
This study has been carried out as part of the “Traceability of origin and authenticity of olive oil by combined genomic and metabolomic approaches” project (OLIV-TRACK, www.dsa.unipr.it/foodhealth/oliv-track/index.html) funded by the European Union (contract QLK1-CT-2002-02386, Quality of Life and Management of Living Resources). The partners of the OLIV-TRACK project are acknowledged for sharing their samples and expertise. We thank MIUR for partial financial support (FIRB “MICRAM”, RBNE01ZB7A and FIRB2003, RBLA03ER38_004). Part of this work was supported by the EU-FPV Marie Curie Training Site scheme. Our thanks to NIAB (Cambridge, UK) for their hospitality and scientific support, to Maria Vurchio for help with administrative issues and to Giada Caredda for the support in the experimental phase.
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