Food allergens are a well acknowledged issue in food industry and are regulated by legislation. The presence of allergens can either origin from the raw material or due to contamination during production. Allergen information on packaging is mandatory although it cannot be accurate in the case of contamination therefore warnings are used. The purpose of the study is the development and validation of a SYBR Green Real Time Polymerase Chain Reaction method using specific primer pairs based on Jug r 1, Jug r 3, and Jug r 4 allergen-coding sequences to improve the sensitivity of Real Time Polymerase Chain Reaction techniques for detection of walnut and almond traces in commercial food products and its comparison with ELISA methodology in terms of detection ability. A total of 100 samples were collected from local markets and were analyzed by Real Time Polymerase Chain Reaction (RT-PCR) and ELISA methods. The results indicated that 16 samples (16%) were found positive in walnut traces and 18 samples (18%) were found positive in almond traces by Real Time Polymerase Chain Reaction of which Elisa identified 14 samples (14%) positive in walnut traces and 15 samples (15%) positive in almond traces. Among them, 4 samples (25%) that contained walnut traces and 6 samples (33.3%) that contained almond traces had no allergen declaration on their label. The improved accuracy of Real Time Polymerase Chain Reaction underlines the importance of this method for allergen detection and quantification in the food industry
The objective of the study is the assessment of the microbial ecology and safety of fish in Greece using next-generation sequencing (NGS) and the correlation of the species of microbial flora with the production of histamine. Fourteen different fish species were obtained from local fish stores (Greece) within 1 day from capture. The initial microbiota in fish flesh was determined using NGS. The main pathogenic bacterial species identified in the tested fish samples included Vibrio spp., Clostridium spp., Staphylococcus, Flavobacterium and Janthinobacterium representing both native freshwater habitats and contaminants arising from different sources, including sewage and direct contamination by wild animals, livestock, and feed. The initial spoilage microbiota of fish consisted of several psychrotrophic Gram-negative bacteria, such as Pseudomonas, Acinetobacter, Moraxella, Shewanella, Psychrobacter, Lactobacillus, Brochothrix and Photobacterium. The results of the study show the potential of the application and the usefulness of NGS for the determination of microbial flora associated with food-borne diseases and spoilage in fish products. Histamine formation correlated with the valid reads (concentration and number of bacteria) and slightly with the genus of the identified microorganisms.
The objective of the study was to determine qualitatively by validated Real Time PCR method the occurrence of genetically modified maize and soybean in commercial food products from the Greek market. 70 independent samples were collected, including products from different categories (i.e. cereal based, biscuits and snacks) which declared either corn or soybean on the labelling. The result of the study indicated that 37.1% of maize and soy products (n=70) displayed in the Greek market have detectable levels of genetically modified maize or soy. These products were identified by specific primers and included common GMΟ detection primers for 35S and NOS terminator. Adequate repeatability and reproducibility was demonstrated for the applied Real Time PCR method, as evaluated by intra- and inter-laboratory tests.
The aim of the present study was the implementation of molecular techniques in the detection and quantification of allergic substances of peanut in various kinds of food products, e.g., breakfast cereals, chocolates and biscuits that are frequently related to allergies. In some cases, the presence of peanuts can be due to contamination during production and are not declared on the label. A total of 152 samples were collected from supermarkets and were analysed by a Real Time PCR method. The results indicated that 125 samples (83,3%) were found positive in peanut traces but the most important finding is that from the 84 samples that had no allergen declaration for peanuts, 48 (57,1%) of them were found positive. In conclusion, Real Time PCR can be a very important tool for the rapid detection and quantification of food allergens.
The purpose of this study was to investigate the possible presence of sesame in commercial foods normally carrying no warning for the allergen, but which may have been subjected to contamination during processing. One hundred units of widely consumed goods with high potential to contain allergenic substances deriving from nuts were analyzed, using sensitive and capable PCR (C-PCR) and Real Time PCR (RT-PCR) methodologies. Of the products examined, 15 (15.0%) declared the presence of sesame, 36 (36.0%) carried no food allergy label, 44 (44.0%) were marked by the phrase "may contain traces of nuts" and 5 (5.0%) carried the indication "may contain sesame traces". The sesame-positive products detected using the C-PCR method were 15 (100%), 12 (33.3%), 14 (31.8%) and 3 (60%), respectively. Using the RT-PCR technique, positive results were obtained for 15 (100%), 18 (50.0%), 18 (20.5%) and 5 (100%) samples, respectively. The results indicate that the PCR methods applied are highly sensitive and selective, which makes them suitable for the detection of sesame traces in food samples. In addition, they can be useful for monitoring the effectiveness of cleaning processes in the production units of the food industry.
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