“…2 Schematic representation of DNA amplification by real time PCR Fig. 3 Amplification plot of tenfold serial-dilution of genetically modified maize Mon810 DNA using ABI Prism® 7900 SDS thermocycler (ABI) (adapted from Chaouachi et al, 2014b ). The figure showed the threshold line which indicates the highest possible noise.…”
Analyses to ensure food safety and quality are more relevant now because of rapid changes in the quantity, diversity and mobility of food. Food-contamination must be determined to maintain health and up-hold laws, as well as for ethical and cultural concerns. Real-time polymerase chain reaction (RT-PCR), a rapid and inexpensive quantitative method to detect the presence of targeted DNA-segments in samples, helps in determining both accidental and intentional adulterations of foods by biological contaminants. This review presents recent developments in theory, techniques, and applications of RT-PCR in food analyses, RT-PCR addresses the limitations of traditional food analyses in terms of sensitivity, range of analytes, multiplexing ability, cost, time, and pointof-care applications. A range of targets, including species of plants or animals which are used as food ingredients, food-borne bacteria or viruses, genetically modified organisms, and allergens, even in highly processed foods can be identified by RT-PCR, even at very low concentrations. Microfluidic RT-PCR eliminates the separate sample-processing step to create opportunities for point-of-care analyses. We also cover the challenges related to using RT-PCR for food analyses, such as the need to further improve sample handling.
“…2 Schematic representation of DNA amplification by real time PCR Fig. 3 Amplification plot of tenfold serial-dilution of genetically modified maize Mon810 DNA using ABI Prism® 7900 SDS thermocycler (ABI) (adapted from Chaouachi et al, 2014b ). The figure showed the threshold line which indicates the highest possible noise.…”
Analyses to ensure food safety and quality are more relevant now because of rapid changes in the quantity, diversity and mobility of food. Food-contamination must be determined to maintain health and up-hold laws, as well as for ethical and cultural concerns. Real-time polymerase chain reaction (RT-PCR), a rapid and inexpensive quantitative method to detect the presence of targeted DNA-segments in samples, helps in determining both accidental and intentional adulterations of foods by biological contaminants. This review presents recent developments in theory, techniques, and applications of RT-PCR in food analyses, RT-PCR addresses the limitations of traditional food analyses in terms of sensitivity, range of analytes, multiplexing ability, cost, time, and pointof-care applications. A range of targets, including species of plants or animals which are used as food ingredients, food-borne bacteria or viruses, genetically modified organisms, and allergens, even in highly processed foods can be identified by RT-PCR, even at very low concentrations. Microfluidic RT-PCR eliminates the separate sample-processing step to create opportunities for point-of-care analyses. We also cover the challenges related to using RT-PCR for food analyses, such as the need to further improve sample handling.
“…Using current quantitative techniques, especially RT-PCR, qPCR, and duplex or multiplex PCR, there were detected different types of unauthorized GMO from maize food (application on 35S promoter) (Bt11, Bt176, MON810 and T25) [14,15] as well as other types of food [16,17]. These methods allow the amplification of multiple target sequences simultaneously, in order to shorten the analysis and monitoring time of food products and to reduce costs of analysis.…”
“…For quantification of MON810 and GA21 in maize, qPCR assays were optimized with limits of detection (LOD) and quantification (LOQ) of 3 and 36 copies respectively 53 . Duplex qPCR method has been developed for identification of four GM maize events, Bt11, Bt176, MON810 and T25 54 . Quadruplex qPCR assay targeting p35S, Tnos and nptII marker gene along with an endogenous gene has been developed for screening of GM tomatoes 17 .…”
Section: Qualitative and Quantitative Pcr Methodsmentioning
Efficient detection strategies for genetically modified (GM) crops need to be in compliance with regulatory frameworks and address consumer concerns. The present review describes widely employed DNA-based technologies for GM detection. Polymerase chain reaction (PCR) and real-time PCR (qPCR) are the methods that can be used for qualitative and quantitative analysis of GM crops due to their specificity, sensitivity and robustness. With increase in number and complexity of genetic elements in newly developed GM events, strategies based on matrix approach, real-time PCR-based multi-target system, loop-mediated isothermal amplification, next generation sequencing, have emerged, which could facilitate cost-effective, rapid, on-site or high throughput GM detection.
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