Detection of the Allergens in Processed Foods Based on Multiple Ligation-dependent Probe Amplification (MLPA)
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Abstract:
In this study, multiplex ligation-dependent probe amplification (MLPA) was applied to simultaneously detect 14 plant-derived allergens from pistachio, Brazil nut, celery, gluten, macadamia nut, sesame, hazelnut, soybean, peanut, sunflower seed, walnut, cashew, almond and mustard. Specific hybridization probes were designed for ITS sequences. After the sample nucleic acid was denatured at 95 ℃, specific binding was carried out with the probes, and target fragments of different sizes were obtained through ligation and PCR amplification reaction. The target fragments were analyzed by capillary electrophoresis to determine whether or not the allergens were present. Using the mixed probe system to detect a single template canonly amplifya single amplification peak, indicating that the probe had high specificity. The detection limit results showed that the lowest detectable DNA concentration of MLPA amplification was 1 ng/μL. Through the detection of 20 actual samples, it is proven that the detection method of allergens in processed foods established in this study on the basis of MLPA has the characteristics of high specificity and high sensitivity, and can be applied to food safety supervision.
