Detection of allergenic substances in foods by a multiplex PCR method

A multiplex PCR (M-PCR) method was developed for the detection of DNAs of plant and three allergenic substances (wheat, buckwheat, and peanut) in foods. Genomic DNAs were extracted from allergenic substances with a commercial ion-exchange type kit. Four primer pairs suitable for the specific detection of plant DNA were designed to establish a M-PCR method detecting simultaneously the specific DNAs of plant and allergenic substances. Our four designed primer pairs and the primer pair described in the Japanese official method were applied to the specific detection of plant DNA. A primer pair of Plant01-5' and Plant01-3' (amplicon size; 161 bp) was the most suitable for the specific detection of plant DNA. M-PCR was performed to detect the specific DNAs of allergenic substances using four primer pairs, a pair of Plant01-5' and Plant01-3', and three pairs for allergenic components described in the Japanese official method. The four specific PCR bands were simultaneously amplified from genomic DNAs of allergenic substances. The proposed method is simple, rapid and inexpensive.