Identification of Mammalian and Poultry Species in Food and Pet Food Samples Using 16S rDNA Metabarcoding

The substitution of more appreciated animal species by animal species of lower commercial value is a common type of meat product adulteration. DNA metabarcoding, the combination of DNA barcoding with next-generation sequencing (NGS), plays an increasing role in food authentication. In the present study, we investigated the applicability of a DNA metabarcoding method for routine analysis of mammalian and poultry species in food and pet food products. We analyzed a total of 104 samples (25 reference samples, 56 food products and 23 pet food products) by DNA metabarcoding and by using a commercial DNA array and/or by real-time PCR. The qualitative and quantitative results obtained by the DNA metabarcoding method were in line with those obtained by PCR. Results from the independent analysis of a subset of seven reference samples in two laboratories demonstrate the robustness and reproducibility of the DNA metabarcoding method. DNA metabarcoding is particularly suitable for detecting unexpected species ignored by targeted methods such as real-time PCR and can also be an attractive alternative with respect to the expenses as indicated by current data from the cost accounting of the AGES laboratory. Our results for the commercial samples show that in addition to food products, DNA metabarcoding is particularly applicable to pet food products, which frequently contain multiple animal species and are also highly prone to adulteration as indicated by the high portion of analyzed pet food products containing undeclared species.

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Detection of Peanut Allergen Traces with a Real Time PCR Assay - The Challenge to Protect Food-Allergic Consumers

Journal of Food Research ◽

10.5539/jfr.v7n1p32 ◽

2017 ◽

Vol 7 (1) ◽

pp. 32 ◽

Cited By ~ 2

Author(s):

Dimitra Houhoula ◽

Stamatios Koussissis ◽

Vladimiros Lougovois ◽

John Tsaknis ◽

Dimitra Kassavita ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Rapid Detection ◽

Food Products ◽

Molecular Techniques ◽

Food Allergens ◽

Pcr Assay ◽

Peanut Allergen ◽

Pcr Method ◽

Detection And Quantification

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.

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Quantitative real-time PCR assays to detect DNA degradation in soy-based food products

Journal of the Science of Food and Agriculture ◽

10.1002/jsfa.3563 ◽

2009 ◽

Vol 89 (7) ◽

pp. 1137-1144 ◽

Cited By ~ 19

Author(s):

Sarah R Murray ◽

Ruth C Butler ◽

Gail M Timmerman-Vaughan

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Food Products ◽

Dna Degradation ◽

Quantitative Real Time Pcr ◽

Pcr Assays

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A Real-Time PCR Method for the Authentication of Common Cuttlefish (Sepia officinalis) in Food Products

Foods ◽

10.3390/foods9030286 ◽

2020 ◽

Vol 9 (3) ◽

pp. 286 ◽

Cited By ~ 2

Author(s):

Amaya Velasco ◽

Graciela Ramilo-Fernández ◽

Carmen G. Sotelo

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Food Products ◽

Polymerase Chain Reaction Method ◽

Cross Reactivity ◽

Sepia Officinalis ◽

Base Pairs ◽

Mitochondrial Coi ◽

Fresh Frozen ◽

Pcr Method

Cephalopods are very relevant food resources. The common cuttlefish (Sepia officinalis) is highly appreciated by consumers and there is a lack of rapid methods for its authentication in food products. We introduce a new minor groove binding (MGB) TaqMan real-time PCR (Polymerase Chain Reaction) method for the authentication of S. officinalis in food products to amplify a 122 base pairs (bp) fragment of the mitochondrial COI (Cytochrome Oxidase I) region. Reference and commercial samples of S. officinalis showed a threshold cycle (Ct) mean of 14.40, while the rest of the species examined did not amplify, or showed a significantly different Ct (p < 0.001). The calculated efficiency of the system was 101%, and the minimum DNA quantity detected was 10−4 ng. No cross-reactivity was detected with any other species, thus, the designed method differentiates S. officinalis from other species of the genus Sepia and other cephalopod species and works for fresh, frozen, grilled, cooked and canned samples of Sepia spp. The method has proved to be reliable and rapid, and it may prove to be a useful tool for the control of fraud in cuttlefish products.

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Application of real-time PCR (qPCR) for characterization of microbial populations and type of milk in dairy food products

Critical Reviews in Food Science and Nutrition ◽

10.1080/10408398.2017.1375893 ◽

2017 ◽

Vol 59 (3) ◽

pp. 423-442 ◽

Cited By ~ 8

Author(s):

Caterina Agrimonti ◽

Benedetta Bottari ◽

Maria Luisa Savo Sardaro ◽

Nelson Marmiroli

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Food Products ◽

Microbial Populations ◽

Dairy Food

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A comparative study of DNA extraction methods to be used in real-time PCR based quantification of ochratoxin A-producing molds in food products

Food Control ◽

10.1016/j.foodcont.2011.12.010 ◽

2012 ◽

Vol 25 (2) ◽

pp. 666-672 ◽

Cited By ~ 23

Author(s):

Alicia Rodríguez ◽

Mar Rodríguez ◽

M. Isabel Luque ◽

Annemarie F. Justesen ◽

Juan J. Córdoba

Keyword(s):

Comparative Study ◽

Real Time ◽

Ochratoxin A ◽

Dna Extraction ◽

Real Time Pcr ◽

Food Products ◽

Extraction Methods ◽

Dna Extraction Methods

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Development of a fast real-time PCR assay based on TaqMan probe for identification of edible rice grasshopper (Oxya chinensis) in processed food products

Food Research International ◽

10.1016/j.foodres.2018.08.059 ◽

2019 ◽

Vol 116 ◽

pp. 441-446 ◽

Cited By ~ 3

Author(s):

Sung-Yeon Kim ◽

Mi-Ju Kim ◽

Seul-Ki Jung ◽

Hae-Yeong Kim

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Food Products ◽

Taqman Probe ◽

Processed Food ◽

Pcr Assay

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Detection of Rendered Meat and Bone Meals by PCR Is Dependent on Animal Species of Origin and DNA Extraction Method

Journal of Food Protection ◽

10.4315/0362-028x-73.6.1090 ◽

2010 ◽

Vol 73 (6) ◽

pp. 1090-1096 ◽

Cited By ~ 9

Author(s):

MICHAEL J. MYERS ◽

DOROTHY E. FARRELL ◽

CHRISTINE M. DEAVER ◽

JACQULINE MASON ◽

HEIDI L. SWAIM ◽

...

Keyword(s):

Real Time ◽

Dna Extraction ◽

Real Time Pcr ◽

Animal Species ◽

Animal Feed ◽

Functional Assay ◽

Threshold Values ◽

Meat And Bone Meal ◽

Bone Meal ◽

Pcr Method

The capability of eight commercially available DNA extraction kits to extract bovine DNA originating in meat and bone meal from fortified feed was evaluated. Four different batches of bovine meat and bone meal (BMBM) were used for DNA extraction with the eight commercial DNA extraction kits. Within each kit, there were minimal differences in the batch-to-batch amounts of extracted DNA. There were differences between the kits in the amounts of DNA that could be extracted from the same amount of starting BMBM. These differences did not translate into differences in the amount of amplifiable DNA from BMBM-fortified dairy feed. Using a validated real-time PCR method, the kit yielding the highest amount extractable DNA was completely unable to yield a positive PCR result; one other kit was also unable to produce a positive PCR result from DNA extracted from BMBM-fortified feed. There was a complete lack of a correlation between the amount of bovine DNA isolated from BMBM by a given extraction kit compared with the relative amounts of DNA isolated from fortified animal feed as evidenced by the cycle threshold values generated using the real-time PCR method. These results demonstrate that extraction of DNA from processed animal protein is different for pure ingredients and fortified animal feeds. These results indicate that a method specifically developed using just animal-derived meat and bone meal may not yield a functional assay when used to detect animal tissues in complete animal feed.

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Detection of Sesame Seed DNA in Foods Using Real-Time PCR

Journal of Food Protection ◽

10.4315/0362-028x-70.4.1033 ◽

2007 ◽

Vol 70 (4) ◽

pp. 1033-1036 ◽

Cited By ~ 27

Author(s):

JENNIFER L. BRZEZINSKI

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Sesame Seed ◽

Food Products ◽

Taqman Probe ◽

2S Albumin ◽

Sesame Seeds ◽

Baked Goods ◽

Pcr Method ◽

Seed Dna

The detection of potentially allergenic foods, such as sesame seeds, in food products is a major concern for the food-processing industry. A real-time PCR method was designed to determine if sesame seed DNA is present in food products. The PCR reaction amplifies a 66-bp fragment of the sesame seed 2S albumin gene, which is detected with a sesame-specific, dual-labeled TaqMan probe. This reaction will not amplify DNA derived from other seeds present in baked goods, such as pumpkin, poppy, and sunflower seeds. Additionally, this assay will not cross-react with DNA from several tree nut species, such as almond, Brazil nut, cashew, hazelnut, and walnut, as well as four varieties of peanut. This assay is sensitive enough to detect 5 pg of purified sesame seed DNA, as well as sesame seed DNA in a spiked wheat cracker sample.

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