Detection of pea in food by real-time polymerase chain reaction (PCR) |
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Authors: | B Brežná L Hudecová T Kuchta |
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Affiliation: | (1) Department of Microbiology and Molecular Biology, Food Research Institute, Priemyselná 4, P.O. Box 25, 842 75 Bratislava 26, Slovakia |
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Abstract: | 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 patés 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. |
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Keywords: | Pea Allergen Polymerase chain reaction (PCR) Meat |
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