Detection of pea in food by real-time polymerase chain reaction (PCR)

• Published in: European food research & technology 2006-03, Vol.222 (5-6), p.600-603
• Main Authors: Brežná, B, Hudecová, L, Kuchta, T
• Format: Article
• Language: English
• Subjects: allergens; Biological and medical sciences; chloroplast DNA; detection; exons; Food; food composition; Food industries; foods; Fruit and vegetable industries; Fundamental and applied biological sciences. Psychology; General aspects; introns; Methods of analysis, processing and quality control, regulation, standards; Microbiology; molecular sequence data; Peas; polymerase chain reaction; protein concentrates; quantitative analysis
• ISSN: 1438-2377; 1438-2385
• DOI: 10.1007/s00217-005-0168-x

A qualitative 5'-nuclease real-time PCR-based method for the detection of pea (Pisum sativum) in food is described. The method consists of DNA isolation by chaotropic solid phase extraction and the subsequent PCR with pea-specific primers and a TaqMan fluorescent probe. The primers and the probe are oriented to the chloroplast DNA intron located between trnL and trnF exons encoding for tRNA. The analytical parameters of the method were inclusivity 100%, exclusivity 100% and the detection limit of 0.11±0.07 ng of pea DNA corresponding to 12±7 diploid pea genome copies. Using a set of model meat pâtés with defined pea contents, a matrix-related detection limit of 0.05% was determined and a linear calibration line was constructed. The presented analytical method was useful for qualitative detection or semiquantitative determination of pea in food products. The method was relatively fast because the analysis could be performed in one working day.