High-Throughput Double Quantitative Competitive Polymerase Chain Reaction for Determination of Genetically Modified Organisms

2005 ◽  
Vol 77 (15) ◽  
pp. 4785-4791 ◽  
Author(s):  
Anastasia K. Mavropoulou ◽  
Theodora Koraki ◽  
Penelope C. Ioannou ◽  
Theodore K. Christopoulos
Keyword(s):  
Polymerase Chain Reaction ◽  
High Throughput ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Chain Reaction ◽  
Competitive Polymerase Chain Reaction ◽  
Polymerase Chain

scholarly journals Applicability of Three Alternative Instruments for Food Authenticity Analysis:GMO Identification

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
A. Burrell ◽  
C. Foy ◽  
M. Burns
Keyword(s):  
Polymerase Chain Reaction ◽  
Gel Electrophoresis ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Fitness For Purpose ◽  
Chain Reaction ◽  
Food Authenticity ◽  
Pcr Products ◽  
Polymerase Chain

Ensuring foods are correctly labelled for ingredients derived from genetically modified organisms (GMOs) is an issue facing manufacturers, retailers, and enforcement agencies. DNA approaches for the determination of food authenticitys often use the polymerase chain reaction (PCR), and PCR products can be detected using capillary or gel electrophoresis. This study examines the fitness for purpose of the application of three laboratory electrophoresis instruments (Agilent Bioanalyzer 2100, Lab901 TapeStation, and Shimadzu MCE-202 MultiNA) for the detection of GMOs using PCR based on a previously validated protocol. Whilst minor differences in the performance characteristics of bias and precision were observed, all three instruments demonstrated their applicability in using this protocol for screening of GMO ingredients.

scholarly journals Detection of genetically modified plants using LAMP (loop-mediated amplification) technologies

2021 ◽  
Vol 17 (1) ◽  
pp. 51-59
Author(s):  
B. V. Sorochynskyi
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Genetically Modified Plants ◽  
Chain Reaction ◽  
Plant Materials ◽  
Advantages And Disadvantages ◽  
Current State ◽  
Polymerase Chain

Purpose. Analysis of the current state and experience on the loop-mediated amplification (LAMP) use to detect genetically modified plants. Methods. Literature search and analysis. Results. General information on the current state and use of the genetically modified plants is provided. Despite the wide distribution of genetically modified plants, the attitude towards them in society continues to remain somewhat wary. About 50 countries have introduced mandatory labeling of GM feed and products, provided that their content exceeds a certain threshold. In order to meet labeling requirements, effective and sensitive methods for detecting known genetic modifications in a variety of plant materials, food products and animal feed must be developed and standardized. The most common approaches to the detection of genetically modified organisms (GMOs) are the determination of specific proteins synthesized in transgenic plants and the detection of new introduced genes. Methods for the determination of GMOs based on the analysis of nucleic acids are more common, since such methods have greater sensitivity and specificity than the analysis of protein composition. Polymerase chain reaction (PCR) method is the main method of nucleic acid analysis, which is now wide used for the detection of GMOs. Loop-mediated amplification (LAMP), which can occur at a constant temperature and therefore does not require the use of expensive equipment may be an alternative to the PCR. Scientific articles about the use of the loop-mediated amplification (LAMP) for the detection of genetically modified plants were analyzed. Advantages and disadvantages of the polymerase chain reaction and loop-mediated amplification are compared. Conclusions. The main criteria for applying a method of GMO detection analysis are as follow: its sensitivity, time of reaction, availability and ease to use, cost of reagents and equipment, and the possibility for simultaneous detection of many samples.

scholarly journals Quantitation of 35S Promoter in Maize DNA Extracts from Genetically Modified Organisms Using Real-Time Polymerase Chain Reaction, Part 2: Interlaboratory Study

2005 ◽  
Vol 88 (2) ◽  
pp. 558-573 ◽  
Author(s):  
Max Feinberg ◽  
Sophie Fernandez ◽  
Sylvanie Cassard ◽  
Chrystèle Charles-Delobel ◽  
Yves Bertheau ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Interlaboratory Study ◽  
Performance Criteria ◽  
Tolerance Interval ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain

Abstract The European Committee for Standardization (CEN) and the European Network of GMO Working Laboratories have proposed development of a modular strategy for stepwise validation of complex analytical techniques. When applied to the quantitation of genetically modified organisms (GMOs) in food products, the instrumental quantitation step of the technique is separately validated from the DNA extraction step to better control the sources of uncertainty and facilitate the validation of GMO-specific polymerase chain reaction (PCR) tests. This paper presents the results of an interlaboratory study on the quantitation step of the method standardized by CEN for the detection of a regulatory element commonly inserted in GMO maize-based foods. This is focused on the quantitation of P35S promoter through using the quantitative real-time PCR (QRT-PCR). Fifteen French laboratories participated in the interlaboratory study of the P35S quantitation operating procedure on DNA extract samples using either the thermal cycler ABI Prism® 7700 (Applied Biosystems, Foster City, CA) or Light Cycler® (Roche Diagnostics, Indianapolis, IN). Attention was focused on DNA extract samples used to calibrate the method and unknown extract samples. Data were processed according to the recommendations of ISO 5725 standard. Performance criteria, obtained using the robust algorithm, were compared to the classic data processing after rejection of outliers by the Cochran and Grubbs tests. Two laboratories were detected as outliers by the Grubbs test. The robust precision criteria gave values between the classical values estimated before and after rejection of the outliers. Using the robust method, the relative expanded uncertainty by the quantitation method is about 20% for a 1% Bt176 content, whereas it can reach 40% for a 0.1% Bt176. The performances of the quantitation assay are relevant to the application of the European regulation, which has an accepted tolerance interval of about ±50%. These data were fitted to a power model (r2 = 0.96). Thanks to this model, it is possible to propose an estimation of uncertainty of the QRT-PCR quantitation step and an uncertainty budget depending on the analytical conditions.

scholarly journals Detection of Genetic Modification in Cured Tobacco Leaf: Proficiency Testing Using Polymerase Chain Reaction-Based Methods

2006 ◽  
Vol 89 (3) ◽  
pp. 693-707
Author(s):  
Ferruccio Gadani ◽  
Martin Ward ◽  
Sue Black ◽  
Neil Harris ◽  
David McDowell ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Proficiency Testing ◽  
Quantitative Methods ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Cauliflower Mosaic Virus ◽  
Chain Reaction ◽  
Tobacco Leaf ◽  
Qualitative And Quantitative ◽  
Polymerase Chain

Abstract The Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA; Paris, France) Task Force Genetically Modified TobaccoDetection Methods investigated the performance of qualitative and quantitative methods based on the polymerase chain reaction (PCR) for the detection and quantitation of genetically modified (GM) tobacco. In the 4 successful rounds of proficiency testing, the cauliflower mosaic virus 35S RNA promoter (CaMV 35S) and the Agrobacterium tumefaciens nopaline synthase terminator (NOS) were selected as target sequences. Blind-coded reference materials containing from 0.1 to 5.0% and from 0.15 to 4% GM tobacco were used in 2 rounds of qualitative and quantitative PCR, respectively. Eighteen laboratories from 10 countries participated in this study. Considering all methods and 2 rounds, the different laboratories were able to detect GM tobacco at the 0.1% level in 46 out of 58 tests in qualitative assays. The results of the proficiency test indicate that both end point screening and real-time quantitative methods are suitable for the detection of genetically modified organisms in tobacco leaf samples having a GM content of 0.1% or higher. The CORESTA proficiency study represents a first step towards the interlaboratory evaluation of accuracy and precision of PCR-based GM tobacco detection, which may lead to the harmonization of analytical procedures and to the enhancement of comparability of testing results produced by different laboratories.

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