DNA-based Methods for Detection of Genetically Modified Events in Food and Supply Chain

Randhawa, Gurinder Jit; Singh, Monika; Sood, Payal

doi:10.18520/cs/v110/i6/1000-1009

Cited by 30 publications

(36 citation statements)

References 71 publications

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“…In each experiment, DNA of the GM maize event was compared with the DNA of unmodified maize. e analytical procedure included several sequential steps: sample preparation (1); thermal treatment (2); genomic DNA extraction (3); DNA assessment by agarose gel electrophoresis and spectrophotometer (4); PCR amplification of endogenous amplicons (5); PCR amplification of GMOspecific amplicons (6); comparison and interpretation of results (7). In this study, we have investigated the effect of different combinations of several important factors on the GMO detection.…”

Section: Resultsmentioning

confidence: 99%

“…e CaMV 35S promoter and Cry1Ab gene is present in both GM maize events Bt176 and MON810 examined in this paper. e transgenic regions of GM plants often contain the CaMV 35S promoter to regulate the transcription of the inserted genes [7]. e Cry1Ab gene of insecticidal protein Cry1Ab δ endotoxin from Bacillus 4 in (a-l)), event Bt176 (lanes 5-8 in (a-c, e-g, i-k)), event MON810 (lanes 5 -8 in (d, h, l)), water (lane 9 in (a-l)), and GelPilot 100 bp ladder (Qiagen) (lane 10 in (a-d)).…”

Section: Impact Of Heat On the Gmo-specific Ampliconsmentioning

confidence: 99%

“…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%

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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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Section: Resultsmentioning

confidence: 99%

Section: Impact Of Heat On the Gmo-specific Ampliconsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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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“…The analytical methods developed for the Bt toxins include DNA‐based methods such as the polymerase chain reaction (PCR), the multiplex polymerase chain reaction (MPCR), the immuno‐PCR technique, and the enzyme‐linked immunosorbent assay (ELISA) . Although the PCR and MPCR techniques can detect the transgenes in the GM plants, use of an immuno‐analytical method, which can detect and quantitate the levels of expression of the transgenic proteins in bulk samples, would be more ideal.…”

Section: Introductionmentioning

confidence: 99%

“…K Rupula et althe multiplex polymerase chain reaction (MPCR), the immuno-PCR technique, and the enzyme-linked immunosorbent assay (ELISA). 15,16 Although the PCR and MPCR techniques can detect the transgenes in the GM plants, use of an immuno-analytical method, which can detect and quantitate the levels of expression of the transgenic proteins in bulk samples, would be more ideal. Given the above considerations, the present study aimed to develop a relatively sensitive immuno-analytical method for the detection and estimation of transgenic Cry1Ac protein.…”

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confidence: 99%

Immuno‐analytical method development for detection of transgenic Cry1Ac protein and its validation

et al. 2019

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BACKGROUND Bacillus thuringiensis (Bt) synthesizes Cry1Ac protein, which is toxic to many lepidopteran pests, and the cry1ac gene has been expressed in several transgenic crop plants. The Cry1Ac protein has been isolated from Bt kurstaki HD73 and purified to homogeneity. Polyclonal antibodies were raised against purified Cry1Ac in rabbits and goat. Sandwich ELISA was developed for Cry1Ac using goat IgG as a coating antibody, and affinity‐purified rabbit IgG as the primary antibody. RESULTS The sensitivity of the assay was in the range of 0.47–1000 ng. It was subsequently employed in validating biological samples. Fifteen different cotton‐seed samples were screened: 12 were found to be Bt positive and 3 Bt negative. The CS7 seeds showed the highest Bt content of 8.51 ± 0.45 μg g−1, followed by CS8 (6.0 ± 0.02 μg g−1), CS15 (5.9 ± 0.03 μg g−1), CS9 (5.5 ± 0.05 μg g−1), and CS10 (4.83 ± 0.013 μg g−1). The CS5 seeds showed Bt content of 3.6 ± 0.21 μg g−1. The F2 generation, CS6 (Kaveri seeds) showed lower Bt content (2.9 ± 0.06 μg g−1). The CL5 samples showed Cry1Ac content of 0.99 ± 0.009 μg g−1. The amount of Cry1Ac protein in leaves, stem, and roots of germinated Bt cotton plants (CS10 and CS4) were 1.76 ± 0.15 μg g−1, 1.9 ± 0.01 μg g−1, 2.0 ± 0.1 μg g−1, and 1.6 ± 0.15 μg g−1, 1.9 ± 0.01 μg g−1, and 2.0 ± 0.01 μg g−1 dry tissue, respectively. CONCLUSION The method developed can be used for screening the expression levels of Cry1Ac in different transgenic Bt cultivars and also spurious Bt cotton seeds procured by farmers. © 2019 Society of Chemical Industry

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Approaches to check unauthorized genetically modified events in supply chain: Challenges and solutions in the Indian context

et al. 2023

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The approval status of a genetically modified (GM) event varies from country to country.GM events approved in one country (considered as authorized GM or AGM) may not necessarily have the same approval status in other countries (considered as unauthorized GM or UGM). Detecting UGM in the supply chain is a challenge as the genetic information is not always available. In India, four Bt cotton events are approved, whereas several other GM events have been imported for research purposes. Many food derivatives (non-GM) are being imported from the countries where GM events of food crops are approved so it is necessary to track the unauthorized entry of GM products. Selected consignments or food products need to be checked for GM status for regulatory compliance. In farmers' fields, the chances of unintentional introgression or adventitious presence of transgenes also need to be monitored in a systematic manner. An appropriate strategy needs to be developed to check for UGM in the food and agricultural supply chain. In this article, approaches for UGM detection have been discussed with a focus on application in the Indian context. Detection methods based on the GMO matrix, multiplex PCR, real-time PCR (qPCR), and loop-mediated isothermal amplification (LAMP) could be employed keeping in view the regulatory requirement or practical application.For checking UGM with unknown genetic construct, methods such as next-generation sequencing (NGS) may be employed. The advantages and disadvantages of the different approaches are discussed in the function of the analytical strategy and its application for control purposes.

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DNA-based Methods for Detection of Genetically Modified Events in Food and Supply Chain

Cited by 30 publications

References 71 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

Immuno‐analytical method development for detection of transgenic Cry1Ac protein and its validation

Approaches to check unauthorized genetically modified events in supply chain: Challenges and solutions in the Indian context

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