Two FAST multiplex real-time PCR reactions to assess the presence of genetically modified organisms in food

Cottenet, Geoffrey; Blancpain, Carine; Sonnard, Véronique; Chuah, Poh Fong

doi:10.1016/j.foodchem.2018.09.050

Cited by 37 publications

(21 citation statements)

References 15 publications

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“…Recently, a new strategy of multiplexing real‐time PCR has been developed, based on the simultaneous detection of six GMO targets exploiting the signal emitted by a single fluorophore (FAM) in a FAST mode. This kind of approach could be very useful, especially to quickly screen samples negative for all the selected targets …”

Section: Discussionmentioning

confidence: 99%

“…This kind of approach could be very useful, especially to quickly screen samples negative for all the selected targets. 19 With the only exception of the p35S/tNOS duplex system, published by Waiblinger et al, all the above mentioned real-time PCR multiplex assays have been developed with the use of master mixes optimized for multiplexing. These reagents are more expensive than *P < 0.05 in the comparison between the duplex and singleplex assay.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, less variable results for the co‐amplification of different targets can be achieved. In recent years, several authors presented methods based on detection and identification of GMO targets using multiplex real‐time PCR . One of the most critical aspects in the development of multiplex PCR methods is the optimization of the experimental conditions with particular regard to the balancing of the components of the reaction mixture.…”

Section: Introductionmentioning

confidence: 99%

“…In recent years, several authors presented methods based on detection and identification of GMO targets using multiplex real-time PCR. [12][13][14][15][16][17][18][19] One of the most critical aspects in the development of multiplex PCR methods is the optimization of the experimental conditions with particular regard to the balancing of the components of the reaction mixture. For this reason almost all of the assays cited above, with the only exception of the p35S/tNOS duplex system described by Waiblinger et al, 12 were based on the use of master mixes optimized for multiplexing.…”

Section: Introductionmentioning

confidence: 99%

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Development and comparative study of a pat/bar real‐time PCR assay for integrating the screening strategy of a GMO testing laboratory

et al. 2020

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BACKGROUND The number and variety of genetically modified organisms (GMOs) used globally for the production of food and feed, and potentially circulating in the European Union (EU), is constantly increasing. This implies an additional effort for the EU enforcement laboratories to optimize available resources, to contain costs and time. A well established approach for streamlining the analytical workflow is the introduction of a screening step, typically based on a smart set of real‐time polymerase chain reaction (PCR) screening methods. The multiplexing strategy, allowing the detection of several screening elements simultaneously, is a further optimization of this step. RESULTS In this study, we present the validation of a real‐time PCR duplex assay for the pat and bar screening elements to be easily incorporated in the GMO diagnostic routine. We also provide a comparison between this method and the related singleplex and pre‐spotted assays. CONCLUSION Our results fully respect all the validation parameters suggested by the Minimum Performance Criteria of the European Network of GMO Laboratories. Furthermore, the duplex assay is equivalent in terms of performance compared to the other two methods, but it shows a higher overall flexibility and cost effectiveness. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development and comparative study of a pat/bar real‐time PCR assay for integrating the screening strategy of a GMO testing laboratory

et al. 2020

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“…DNA diagnostics represent the most e ective tool for the analysis of food ingredients as DNA is the most stable molecule during food processing. Moreover, the DNA-based polymerase chain reaction (PCR) is recognized as a reference method for GMO detection [6][7][8]. However, food production may cause fragmentation of genomic DNA and a ect PCR analysis [9][10][11][12][13][14][15][16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

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

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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.

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Performance assessment of digital PCR for the quantification of GM-maize and GM-soya events

2019

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Two FAST multiplex real-time PCR reactions to assess the presence of genetically modified organisms in food

Cited by 37 publications

References 15 publications

Development and comparative study of a pat/bar real‐time PCR assay for integrating the screening strategy of a GMO testing laboratory

Development and comparative study of a pat/bar real‐time PCR assay for integrating the screening strategy of a GMO testing laboratory

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

Performance assessment of digital PCR for the quantification of GM-maize and GM-soya events

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