Combined effects of matrix and gene marker on the real‐time <scp>PCR</scp> detection of wheat

Martín-Fernández, Begoña; Costa, Joana; Oliveira, M. Beatriz P.P.; López-Ruíz, Beatríz; Mafra, Isabel

doi:10.1111/ijfs.13141

Cited by 10 publications

(4 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…is proves the significant role of the DNA marker in the GMO detection. Our findings coincide with the results of Martin-Fernaandez et al [37]. ey have reported that the sensitivity of the detection of wheat was considerably affected by the marker gene.…”

Section: Impact Of Heat On the Gmo-specific Ampliconssupporting

confidence: 93%

Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize

Bitskinashvili

Gabriadze²,

Kutateladze³

et al. 2019

Journal of Food Quality

View full text Add to dashboard Cite

Reliable detection of genetically modi ed (GM) maize is signi cant for food authenticity, labelling, quality, and safety assessment.is study aims to evaluate the factors in uencing degradation and polymerase chain reaction (PCR) ampli cation of DNA from the wild type and transgenic maize (events Bt-176 and MON810) during thermal treatment at 100°C and 121°C. A new PCR method was developed targeting the Cry1Ab gene to detect insect-resistant GM plants. e yield of genomic DNAs extracted by the DNeasy plant mini kit dramatically decreased while DNAs obtained by cetyltrimethyl ammonium bromide-(CTAB-) based method did not show any visible changes in the yield by the time of processing. Treatment at 100°C did not signi cantly a ect either genomic DNAs or amplicons. Heating at 121°C induced time-dependent degradation of genomic DNAs and exogenous Cry1Ab gene; however, it did not have any considerable in uence on the exogenous 141 bp amplicons or endogenous amplicons in the range of 102 bp to 226 bp with the exception of the event MON810 extracted by the DNeasy plant mini kit. More yield was observed at 226 bp than 140 bp fragment of the invertase gene. e 141 bp fragment of the transgenic CaMV 35S promoter exhibited the highest thermal stability of all the examined amplicons. Analysis of foodstu s demonstrated 102 bp amplicons speci c for the zein gene as the e ective marker to detect maize in the processed foods. e obtained results demonstrate that PCR-based detection of the wild type and transgenic maize is dependent on the combination of di erent parameters of crucial factors such as temperature and duration of exposure, transgenic event, DNA extraction method, DNA marker, and size and location of amplicons.

show abstract

Section: Impact Of Heat On the Gmo-specific Ampliconssupporting

confidence: 93%

Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize

Bitskinashvili

Gabriadze²,

Kutateladze³

et al. 2019

Journal of Food Quality

View full text Add to dashboard Cite

show abstract

“…Moreover, the LOD of raw sausages is 0.005% (50 mg/kg, w/w) with optimal PCR performance parameters (R 2 = 0.990, slope = −3.122, and PCR efficiency = 109.1%), contrarily to raw ham that achieved a LOD of 0.01% (100 mg/kg, w/w) and a correlation coefficient of 0.951, below the acceptable value of 0.98 (Figure 3c, Table 2). The effect of food matrix in food allergen detection by DNA-based methods has been reported by some authors [18,29,30]. The use of reference materials with different calibration models in order to simulate the real food matrices is extremely important.…”

Section: Effect Of Food Matrix and Thermal Treatmentmentioning

confidence: 99%

Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method

Villa

Costa

2019

Biomolecules

Self Cite

View full text Add to dashboard Cite

Milk ingredients are often included in a wide range of meat products, such as cooked hams and sausages, to improve technological characteristics. However, milk proteins are also important food allergens. The aim of this study was the development of a highly sensitive and specific real-time PCR system targeting the 12S rRNA gene of Bos domesticus for the detection and quantification of milk as an allergenic ingredient in processed meat products. The method was able to achieve an absolute limit of detection (LOD) of 6 fg of milk DNA. Using a normalized approach (∆Ct method) for the detection of milk protein concentrate (MPC), it was possible to obtain sensitivities down to 0.01% (w/w) of MPC in model hams (raw and cooked) and autoclaved sausages, and 0.005% in raw sausage mixtures. The developed systems generally presented acceptable PCR performance parameters, being successfully validated with blind samples, applied to commercial samples, and further compared with an immunochemical assay. Trace amounts of milk material were quantified in two out of 13 samples, but the results mostly infer the excessive practice of the precautionary labeling.

show abstract

“…This technique is highly sensitive (5–50 pg of DNA) and can lead to species identification (useful for cross-contamination studies). Real time PCR methods can be also used for the quantitative detection of gluten when using model mixtures [ 46 , 47 ]. Moreover, PCR methods perform indirect detection (not pathogenic compounds of gluten—rather the DNA that codifies for it) [ 48 ].…”

Section: Methods For Gluten Detection In Food Samplesmentioning

confidence: 99%

From Polyclonal Sera to Recombinant Antibodies: A Review of Immunological Detection of Gluten in Foodstuff

Garcia-Calvo

García-García

Madrid

et al. 2020

Foods

View full text Add to dashboard Cite

Gluten is the ethanol-soluble protein fraction of cereal endosperms like wheat, rye, and barley. It is widely used in the food industry because of the physical–chemical properties it gives to dough. Nevertheless, there are some gluten-related diseases that are presenting increasing prevalences, e.g., celiac disease, for which a strict gluten-free diet is the best treatment. Due to this situation, gluten labeling legislation has been developed in several countries around the world. This article reviews the gluten immune detection systems that have been applied to comply with such regulations. These systems have followed the development of antibody biotechnology, which comprise three major methodologies: polyclonal antibodies, monoclonal antibodies (mAbs) derived from hybridoma cells (some examples are 401.21, R5, G12, and α-20 antibodies), and the most recent methodology of recombinant antibodies. Initially, the main objective was the consecution of new high-affinity antibodies, resulting in low detection and quantification limits that are mainly achieved with the R5 mAb (the gold standard for gluten detection). Increasing knowledge about the causes of gluten-related diseases has increased the complexity of research in this field, with current efforts not only focusing on the development of more specific and sensitive systems for gluten but also the detection of protein motifs related to pathogenicity. New tools based on recombinant antibodies will provide adequate safety and traceability methodologies to meet the increasing market demand for gluten-free products.

show abstract

Combined effects of matrix and gene marker on the real‐time PCR detection of wheat

Cited by 10 publications

References 24 publications

Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize

Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize

Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method

From Polyclonal Sera to Recombinant Antibodies: A Review of Immunological Detection of Gluten in Foodstuff

Contact Info

Product

Resources

About