ESTABLISHING THE REAL-TIME PCR ASSAY TO DETECT TROPOMYOSIN GENE CAUSING ALLERGY OF SEAFOOD ORIGIN

Abstract

Seafood is an important dietary source of protein in the world. In addition to the many benefits of seafood such as heart prevention, micronutrient supplementation, high macronutrients, etc., seafood itself is the source of allergies affecting health, even threatening to human’s life with symptoms related to the immune system: red rash, dermatitis, asthma, etc. Gen Tropomyosin - encodes for the protein Tropomyosin, the main allergenic molecule of seafood origin. Therefore, this study aims to establish a detection process, namely Real-time PCR assay, to detect the Tropomyosin gene - a seafood-derived allergen that can be applied in Vietnam. The results, successfully built a positive control for the Real-time assay of detecting the Tropomyosin gene: recombinant vector pUC19-Tropo; Successful design of Real-time PCR assay with TropoF and TropoR primers (amplification of Tropomyosin gene), N18SF and N18SR primers (amplification of 18S rRNA gene – internal control) applied in Tropomyosin gene amplification of seafood origin with the following parameters: detection limit: 101,% CV of the intra assay: 3.64%,% CV of the inter assay: 2.81%. The percentage of commercial food samples positive for Tropomyosin gene was 37/56, reaching 66.11%. Can the successful detection of an allergen help food manufacturers to put a mark on the label so that consumers can choose to use it? Thereby, it helps to reduce the rate of allergies in people using processed foods.