scholarly journals Exploratory and confirmatory molecular approaches to determine genetically modified status in different crops

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Hanaa Abdel-Sadek Oraby ◽  
Nadia Aboul-Ftooh Aboul-Maaty ◽  
Hayam Ahmad Al-Sharawi
Keyword(s):  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Melting Curve ◽  
Conventional Polymerase Chain Reaction ◽  
Melting Curve Analysis ◽  
Target Sequence ◽  
Molecular Approaches ◽  
Detection And Identification ◽  
Food And Feed ◽  
Detection And Quantification

Abstract Background One of the parameters required for the assessment of food and feed safety is detection and identification of genetically modified organisms. Legislation in some countries necessitates detection and quantification of modification in food and feed samples. Scientists have raised concern about safety of antibiotic resistance marker (ARM) genes used for transformation of crops intended for human and animal consumption. In the present work two molecular approaches have been adopted: one exploratory; for detection and quantification of ARM genes in tested plant samples and the other confirmatory; to determine the specificity/reliability of the obtained results. Results Results revealed that primers for neomycin phosphotransferase (nptII) and aminoglycoside 3″ adenyl-transferase (aadA) were amplified in the majority of the 36 DNA screened samples. Melting curve analysis using hygromycin phosphotransferase (aphIV) gene as target sequence for the fluorescent-based detection approach was performed to ensure reliability and specificity of this procedure and to confirm results obtained by using conventional polymerase chain reaction (PCR). Quantitative RT-PCR results and validation analysis followed, revealed that all of the tested DNA samples were not violating the European legislation for GMOs labeling (0.9%). Conclusions The results fully demonstrated the reproducibility, sensitivity/specificity of the adopted approaches for detection and quantification of even traces of GMO contents. Applying measurement uncertainty (MU) procedures presented in this work will help decision makers to ensure compliance with International Legislation and Regulations. This in its turn will facilitate and enhance trading with countries having compelling labeling regulations.

scholarly journals GMO Genetic Elements Thesaurus (GMO-GET): a controlled vocabulary for the consensus designation of introduced or modified genetic elements in genetically modified organisms

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Paulien Adamse ◽  
Emilie Dagand ◽  
Karen Bohmert-Tatarev ◽  
Daniela Wahler ◽  
Manoela Miranda ◽  
...  
Keyword(s):  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Controlled Vocabulary ◽  
Data Systems ◽  
Genetic Elements ◽  
Related Information ◽  
Detection And Identification ◽  
Unique Approach ◽  
The Subject ◽  
And Control

Abstract Background Various databases on genetically modified organisms (GMOs) exist, all with their specific focus to facilitate access to information needed for, e. g., the assistance in risk assessment, the development of detection and identification strategies or inspection and control activities. Each database has its unique approach towards the subject. Often these databases use different terminology to describe the GMOs. For adequate GMO addressing and identification and exchange of GMO-related information it is necessary to use commonly agreed upon concepts and terminology. Result A hierarchically structured controlled vocabulary describing the genetic elements inserted into conventional GMOs, and GMOs developed by the use of gen(om)e-editing is presented: the GMO genetic element thesaurus (GMO-GET). GMO-GET can be used for GMO-related documentation, including GMO-related databases. It has initially been developed on the basis of two GMO databases, i.e. the Biosafety Clearing-House and the EUginius database. Conclusion The use of GMO-GET will enable consistent and compatible information (harmonisation), also allowing an accurate exchange of information between the different data systems and thereby facilitating their interoperability. GMO-GET can also be used to describe genetic elements that are altered in organisms obtained through current targeted genome-editing techniques.

scholarly journals Calculation of Measurement Uncertainty in Quantitative Analysis of Genetically Modified Organisms Using Intermediate PrecisionA Practical Approach

2007 ◽  
Vol 90 (2) ◽  
pp. 582-586 ◽  
Author(s):  
Jana Žel ◽  
Kristina Gruden ◽  
Katarina Cankar ◽  
Dejan tebih ◽  
Andrej Blejec
Keyword(s):  
Nucleic Acids ◽  
Measurement Uncertainty ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Laboratory Diagnostics ◽  
Quantitative Characterization ◽  
Intermediate Precision ◽  
Food And Feed ◽  
Repeatability And Reproducibility

Abstract Quantitative characterization of nucleic acids is becoming a frequently used method in routine analysis of biological samples, one use being the detection of genetically modified organisms (GMOs). Measurement uncertainty is an important factor to be considered in these analyses, especially where precise thresholds are set in regulations. Intermediate precision, defined as a measure between repeatability and reproducibility, is a parameter describing the real situation in laboratories dealing with quantitative aspects of molecular biology methods. In this paper, we describe the top-down approach to calculating measurement uncertainty, using intermediate precision, in routine GMO testing of food and feed samples. We illustrate its practicability in defining compliance of results with regulations. The method described is also applicable to other molecular methods for a variety of laboratory diagnostics where quantitative characterization of nucleic acids is needed.

scholarly journals Log transformation of proficiency testing data on the content of genetically modified organisms in food and feed samples: is it justified?

2019 ◽  
Vol 412 (5) ◽  
pp. 1129-1136 ◽  
Author(s):  
Wim Broothaerts ◽  
Fernando Cordeiro ◽  
Philippe Corbisier ◽  
Piotr Robouch ◽  
Hendrik Emons
Keyword(s):  
Normal Distribution ◽  
Proficiency Testing ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Data Transformation ◽  
Evaluation Procedure ◽  
Reference Laboratory ◽  
Log Data ◽  
Log Transformation ◽  
Food And Feed

AbstractThe outcome of proficiency tests (PTs) is influenced, among others, by the evaluation procedure chosen by the PT provider. In particular for PTs on GMO testing a log-data transformation is often applied to fit skewed data distributions into a normal distribution. The study presented here has challenged this commonly applied approach. The 56 data populations from proficiency testing rounds organised since 2010 by the European Union Reference Laboratory for Genetically Modified Food and Feed (EURL GMFF) were used to investigate the assumption of a normal distribution of reported results within a PT. Statistical evaluation of the data distributions, composed of 3178 reported results, revealed that 41 of the 56 datasets showed indeed a normal distribution. For 10 datasets, the deviation from normality was not statistically significant at the raw or log scale, indicating that the normality assumption cannot be rejected. The normality of the five remaining datasets was statistically significant after log-data transformation. These datasets, however, appeared to be multimodal as a result of technical/experimental issues with the applied methods. On the basis of the real datasets analysed herein, it is concluded that the log transformation of reported data in proficiency testing rounds is often not necessary and should be cautiously applied. It is further shown that the log-data transformation, when applied to PT results, favours the positive performance scoring for overestimated results and strongly penalises underestimated results. The evaluation of the participants’ performance without prior transformation of their results may highlight rather than hide relevant underlying analytical problems and is recommended as an outcome of this study.

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