Development and validation of a SYBR-Green I real-time PCR protocol to detect hazelnut (Corylus avellana L.) in foods through calibration via plasmid reference standard.
Many tree nuts are considered to be a serious problem in food safety, because of the presence of causative factors in IgE-mediated food allergies. Among these, hazelnut (Corylus avellana L.) seeds are largely used in a range of confectionery products and contain many well-characterized allergens. DNA-based methods and ELISA tests may prove to be useful to assess the presence of hidden ingredients in foods. The aim of this work was the development and validation of a species-specific SYBR Green I real-time PCR protocol for the detection of hazelnut in foods. A novel efficient primer pair on the Cor a 8 genomic coding region was designed by preparing a plasmid vector-based internal reference standard to calibrate the PCR. A good sensitivity, down to 20 (genomic) and 15 (plasmid) DNA copies, was established. All of the commercial samples considered in our study (containing hazelnut as ingredient or as a potential trace cross-contamination) were effectively amplified by PCR, showing a perfect correspondence with an ELISA commercial test, employed as a reference standard method.
D'Andrea
Matteo
M
DISCAFF and DFB Center, Universita del Piemonte Orientale A. Avogadro, Novara, Italy.
Coïsson
Jean Daniel
JD
Travaglia
Fabiano
F
Garino
Cristiano
C
Arlorio
Marco
M
D052179:Antigens, Plant / Q000235:genetics* / Q000276:immunology; D052160:Benzothiazoles; D002138:Calibration; D031211:Corylus / Q000235:genetics* / Q000276:immunology; D003959:Diamines; D005456:Fluorescent Dyes / Q000737:chemistry; D009930:Organic Chemicals / Q000737:chemistry; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011804:Quinolines; D012015:Reference Standards; D012680:Sensitivity and Specificity
D052179:Antigens, Plant; D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D010940:Plant Proteins; D011804:Quinolines; C098022:SYBR Green I