New Multiplexing Tools for Reliable GMO Detection

Pla, María; Nadal, Anna; Baeten, Vincent; Bahrdt, C.; Berben, Gilbert; Bertheau, Yves; Coll, Anna; Dijk, Jeroen P. van; Dobnik, David; Fernández-Pierna, Juan A.; Gruden, Kristina; Hamels, Sandrine; Holck, Askild Lorentz; Holst‐Jensen, Arne; Janssen, Éric; Kok, E.J.; Paz, José Luís La; Laval, Valérie; Leimanis, Serge; Malcevschi, Alessio; Marmiroli, Nelson; Morisset, Dany; Prins, T.W.; Remacle, José; Ujhelyi, Gabriella; Wulff, Doerte

doi:10.1002/9781118373781.ch19

Cited by 6 publications

(7 citation statements)

References 78 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Multiplex PCR techniques have been applied for qualitative detection and identification as well as for quantification of GMOs (Hohne et al, 2002 ; Kim et al, 2006 ; Nikolić et al, 2008 ; Waiblinger et al, 2008 ; Samson et al, 2013 ). A number of new approaches (Querci et al, 2010 ; Pla et al, 2012 ) have been developed that involve the use of PCR with multiple targets and consecutive detection and identification of the amplification products using capillary gel electrophoresis (CGE) (Heide et al, 2008 ; Nadal et al, 2009 ; Holck et al, 2010 ), hybridization in microarrays (Leimanis et al, 2006 ; Hamels et al, 2009 ; Li et al, 2015 ), and next generation sequencing (Holst-Jensen et al, 2012 ; Milavec et al, 2014 ). The cost-efficiency and high-throughput could also be achieved by the combinatory SYBR Green qPCR and matrix-based approach (Chaouachi et al, 2008 ; Holst-Jensen, 2009 ; Querci et al, 2009 ; Van den Bulcke et al, 2010 ; Barbau-Piednoir et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

Datukishvili

Kutateladze²,

Gabriadze³

et al. 2015

Front. Microbiol.

View full text Add to dashboard Cite

We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs). New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV) 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS) terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps) gene and 258 bp fragment of Cry1Ab delta-endotoxin (cry1Ab) gene for GMO screening. The certified reference materials containing Roundup Ready soybean (RRS) and maize MON 810 were applied for the development and optimization of uniplex and multiplex PCR systems. Evaluation of amplification products by agarose gel electrophoresis using negative and positive controls confirmed high specificity and sensitivity at 0.1% GMO for both RRS and MON 810. The fourplex PCR was developed and optimized that allows simultaneous detection of three common transgenic elements, such as: CaMV 35S promoter, NOS terminator, epsps gene together with soybean-specific lectin gene. The triplex PCR developed enables simultaneous identification of transgenic elements, such as: 35S promoter and cry1Ab gene together with maize zein gene. The analysis of different processed foods demonstrated that multiplex PCR methods developed in this study are useful for accurate and fast screening of GM food products.

show abstract

Section: Introductionmentioning

confidence: 99%

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

Datukishvili

Kutateladze²,

Gabriadze³

et al. 2015

Front. Microbiol.

View full text Add to dashboard Cite

show abstract

“…Methods, selected for evaluation and comparison (assessment) in this study, are listed in Table 1. Four of the methods represent the qPCR system currently used in routine GMO diagnostics covering different applications: simplex and multiplex screening/identification (Alary et al 2002;Kuribara et al 2002;Pla et al 2013) and simplex quantification (Holck et al 2002). Two additional qPCR applications (SIMQUANT; Berdal et al 2008) use qPCR chemistry together with the limiting dilutions principle, which is near to the idea of the ddPCR-based methods, of which two were included Dobnik et al 2015).…”

Section: Analytical Methods Assessed In This Studymentioning

confidence: 99%

Decision Support for the Comparative Evaluation and Selection of Analytical Methods: Detection of Genetically Modified Organisms as an Example

et al. 2018

View full text Add to dashboard Cite

The selection of the best-fit-for-purpose analytical method to be implemented in the laboratory is difficult due to availability of multiple methods, targets, aims of detection, and different kinds and sources of more or less reliable information. Several factors, such as method performance, practicability, cost of setup, and running costs need to be considered together with personnel training when selecting the most appropriate method. The aim of our work was to prepare a flexible multicriteria decision analysis model suitable for evaluation and comparison of analytical methods used for the purpose of detecting and/or quantifying genetically modified organisms, and to use this model to evaluate a variety of changing analytical methods. Our study included selection of PCR-, isothermal-, protein-, microarray-, and next-generation sequencing-based methods in simplex and/or multiplex formats. We show that the overall result of their fitness for purpose is relatively similar; however, individual criteria or a group of related criteria exposed more substantial differences between the methods. The proposed model of this decision support system enables easy modifications and is thus suitable for any other application of complex analytical methods.

show abstract

“…In a second step, the PCR products are analyzed using the CGE, microarray, or Luminex platforms. Despite the fact that these technologies present a higher throughput than qPCR, their multiplexing level is still influenced by the inherent properties of PCR which limit the number of reactions at commonly ten targets per PCR assay [ 133 , 134 ].…”

Section: Gmo Detection Approachesmentioning

confidence: 99%

“…With the microarray technology applied to GMO detection, GM targets are amplified by PCR, using target-specific and/or universal primers, prior to being hybridized on the array, allowing the simultaneous detection of more than 250 000 targets in one assay ( Figure 1 and Table 5 ) [ 136 ]. Compared to the qPCR, the microarray strategy presents thus a well higher throughput but a slightly weaker sensitivity [ 133 , 137 ]. One approach, called multiplex quantitative DNA array-based PCR (MQDA-PCR), tested on transgenic maize events, consists of a first PCR using target-specific primers that harbor a universal tail allowing using universal primers in the second PCR.…”

Section: Gmo Detection Approachesmentioning

confidence: 99%

“…The performance of the DualChip GMO system, targeting fourteen elements, was also validated through an EU collaborative ring trial. An upgraded version of this system (DualChip GMO V2.0) presents a higher GMO coverage in targeting thirty elements ( Table 5 ) [ 59 – 61 , 133 , 138 ]. Most recently, a multiplex amplification on a chip with readout on an oligo microarray (MACRO) system, targeting ninety-one targets to cover a broad spectrum of GMO, was also reported [ 139 ].…”

Section: Gmo Detection Approachesmentioning

confidence: 99%

See 1 more Smart Citation

Current and New Approaches in GMO Detection: Challenges and Solutions

Fraiture

Herman

Taverniers³

et al. 2015

BioMed Research International

117

View full text Add to dashboard Cite

In many countries, genetically modified organisms (GMO) legislations have been established in order to guarantee the traceability of food/feed products on the market and to protect the consumer freedom of choice. Therefore, several GMO detection strategies, mainly based on DNA, have been developed to implement these legislations. Due to its numerous advantages, the quantitative PCR (qPCR) is the method of choice for the enforcement laboratories in GMO routine analysis. However, given the increasing number and diversity of GMO developed and put on the market around the world, some technical hurdles could be encountered with the qPCR technology, mainly owing to its inherent properties. To address these challenges, alternative GMO detection methods have been developed, allowing faster detections of single GM target (e.g., loop-mediated isothermal amplification), simultaneous detections of multiple GM targets (e.g., PCR capillary gel electrophoresis, microarray, and Luminex), more accurate quantification of GM targets (e.g., digital PCR), or characterization of partially known (e.g., DNA walking and Next Generation Sequencing (NGS)) or unknown (e.g., NGS) GMO. The benefits and drawbacks of these methods are discussed in this review.

show abstract

New Multiplexing Tools for Reliable GMO Detection

Cited by 6 publications

References 78 publications

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

Decision Support for the Comparative Evaluation and Selection of Analytical Methods: Detection of Genetically Modified Organisms as an Example

Current and New Approaches in GMO Detection: Challenges and Solutions

Contact Info

Product

Resources

About