Detection and Quantitation of Genetically Modified Maize (Bt-176 Transgenic Maize) by Applying Ligation Detection Reaction and Universal Array Technology

Bordoni, Roberta; Mezzelani, Alessandra; Consolandi, Clarissa; Frosini, Andrea; Rizzi, Ermanno; Castiglioni, Bianca; Salati, Claudia; Marmiroli, Nelson; Marchelli, Rosangela; Bernardi, L.; Battaglia, Cristina; Bellis, Gianluca De

doi:10.1021/jf034871e

Cited by 39 publications

(19 citation statements)

References 12 publications

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“…In comparison to other array-based GMO detection methods, the method presented in this study shows comparable performances in terms of specificity and relative sensitivity; most other methods being able to detect as low as 0.1% GM contents (6,9,29,31), and some being slightly less sensitive (8,10). The absolute sensitivity of the NAIMA amplification method combined with microarray detection (∼60 pg) is also favourable when compared to the above-cited methods, since several methods require starting amounts of DNA from 10 to 500 ng (8,10,31), two methods requiring 60 pg (29) and 300 pg (6).…”

Section: Resultsmentioning

confidence: 81%

“…Regarding the performance, NAIMA amplification shows very fast kinetics that are comparable with the fastest qPCR systems currently available, as the duration of a qPCR amplification run varies from 30 min to 2 h (28). NAIMA is also generally a faster amplification method when compared to other multiplex amplification methods combined with microarray detection applied to GMO diagnostics (6–10,12,29). Most of the qPCR methods validated by the CRL for GMO detection include cycles of 75 s, for which a maximum of 7·10 10 -fold amplification rate (for 36 cycles) is possible in 45 min at 100% reaction efficiency (http://gmo-crl.jrc.it/statusofdoss.htm).…”

Section: Resultsmentioning

confidence: 99%

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NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs

Morisset

Dobnik

Hamels

et al. 2008

Nucleic Acids Research

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We have developed a novel multiplex quantitative DNA-based target amplification method suitable for sensitive, specific and quantitative detection on microarray. This new method named NASBA Implemented Microarray Analysis (NAIMA) was applied to GMO detection in food and feed, but its application can be extended to all fields of biology requiring simultaneous detection of low copy number DNA targets. In a first step, the use of tailed primers allows the multiplex synthesis of template DNAs in a primer extension reaction. A second step of the procedure consists of transcription-based amplification using universal primers. The cRNA product is further on directly ligated to fluorescent dyes labelled 3DNA dendrimers allowing signal amplification and hybridized without further purification on an oligonucleotide probe-based microarray for multiplex detection. Two triplex systems have been applied to test maize samples containing several transgenic lines, and NAIMA has shown to be sensitive down to two target copies and to provide quantitative data on the transgenic contents in a range of 0.1–25%. Performances of NAIMA are comparable to singleplex quantitative real-time PCR. In addition, NAIMA amplification is faster since 20 min are sufficient to achieve full amplification.

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Section: Resultsmentioning

confidence: 81%

Section: Resultsmentioning

confidence: 99%

NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs

Morisset

Dobnik

Hamels

et al. 2008

Nucleic Acids Research

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“…Bordoni et al (2004) reported a sensitivity of 0.1% for the Bt176 transgenic maize and can identify the presence of 0.5% transgenic events within complex mixtures of RRS, Bt11, MON810, GA21 and Bt176 (Bordoni et al, 2005). Recently, a peptide nucleic acid (PNA) array platform was reported for the detection of GMO in food: in this paper , PNA array showed a high selectivity for several 5% GM events and two specific plant genes (lectin for soybean and zein for maize).…”

Section: Discussionmentioning

confidence: 99%

“…Different DNA microarray approaches have been developed to be used in combination with multiplex PCRs: a multiplex DNA array-based PCR allowing quantification of transgenic maize in food and feed (Rudi et al, 2003); a ligation detection reaction coupled with an universal array technology allows the detection of the Bt176 transgenic maize (Bordoni et al, 2004) or five transgenic events (Bordoni et al, 2005); and recently, a peptide nucleic acid array approach was developed for the detection of five transgenic events and two plant species . These methods used fluorescent probes, which require costly material and are photosensitive, thus limiting the common use of microarrays for GM detection.…”

Section: Introductionmentioning

confidence: 99%

A Microarray-based Detection System for Genetically Modified (GM) Food Ingredients

Leimanis

Hernández

Fernandez³

et al. 2006

Plant Mol Biol

119

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A multiplex DNA microarray chip was developed for simultaneous identification of nine genetically modified organisms (GMOs), five plant species and three GMO screening elements, i.e. the 35S promoter, the nos terminator and the nptII gene. The chips also include several controls, such as that for the possible presence of CaMV. The on-chip detection was performed directly with PCR amplified products. Particular emphasis was placed on the reduction of the number of PCR reactions required and on the number of primers present per amplification tube. The targets were biotin labelled and the arrays were detected using a colorimetric methodology. Specificity was provided by specific capture probes designed for each GMO and for the common screening elements. The sensitivity of the assay was tested by experiments carried out in five different laboratories. The limit of detection was lower than 0.3% GMO for all tests and in general around 0.1% for most GMOs. The chip detection system complies with the requirements of current EU regulations and other countries where thresholds are established for the labelling of GMO.

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“…12 The ligation detection reaction was combined with a universal array approach to detect and quantitate the PCR-amplified Cry1A(b) gene from BT-176 transgenic maize with excellent specificity and high sensitivity. 13 An immono-PCR based method showed minimum detection limit 21.6-436 ng to detect insecticidal protein Cry1Ac toxin, produced by Bacillus thuringiensis. 14 In a need to apply a screening method that is sensitive and unambiguous in identifying the different transformation events, a small amount of DNA required by TAIL-PCR was easily recovered from small transformant that allows rapid verification of T-DNA integration and detection of separate gene transfer event.…”

Section: Tracking Of Genetically Engineered Dna In the Environmentmentioning

confidence: 99%

Regulation and Safety Assessment of Genetically Engineered Food

Singh

2010

Studies in Ethics, Law, and Technology

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Transgenic technologies avails new ways that alter plants and animals to be better suited for applications in food, feed, and processing. The ability to express foreign genes and proteins opens the door to producing many commercially important industrial and pharmaceutical products. However, despite the promise of these technologies, there are many concerns about the environmental impact of genetically engineered (GE) food plants and how to contain them. Risk assessment and monitoring are vital for this industry: the regulatory agencies aimed to monitor the specific environment and public health hazards associated with GE food and organisms. In the United States, the FDA, USDA, and EPA are responsible for these regulations. Several agencies in other countries also monitor GE foods and frame guidelines for the safe application of recombinant genes in agro-industries. This article gives an overview on the tracking of GE DNA in foods and the general public's concerns about them. The role of regulatory agencies are also summarized in regulating GE products while ensuring the public health.

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Detection and Quantitation of Genetically Modified Maize (Bt-176 Transgenic Maize) by Applying Ligation Detection Reaction and Universal Array Technology

Cited by 39 publications

References 12 publications

NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs

NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs

A Microarray-based Detection System for Genetically Modified (GM) Food Ingredients

Regulation and Safety Assessment of Genetically Engineered Food

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