Application of DNA- and Protein-Based Detection Methods in Agricultural Biotechnology

Abstract

DNA-and protein-based detection methods are widely used tools for monitoring biotechnology-derived crops and their products globally. Agricultural biotechnology companies, food/feed suppliers and supply chains, diagnostic testing companies, and regulatory authorities heavily rely on these two technologies for product development, seed production, compliance, and contractual needs. The primary use of DNA-and protein-based detection methods is either to verify the presence or absence of genetically engineered (GE) materials or to quantify the amount of GE material present in a product. This review describes key parameters of DNA-and protein-based detection methods, and thorough assessment of their applications and their advantage and limitations in agricultural biotechnology are discussed in detail. The review highlights the principle and considerations of detection method selection, which will equip users to choose suitable technology and obtain reliable test results. The review also compares the compatibility of the two technologies in GE product testing using a case study. KEYWORDS: genetically modified (GM), genetically modified organism (GMO), genetically engineered (GE), GM food testing, food analysis, DNA detection methods, protein detection methods, Bioengineered (BE)

Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize

“…Due to this evidence, we examined maize-specific and GMO-specific amplicons. e PCR products were selected within the size range between 102 bp and 258 bp, foreseeing the efficiency of short amplicons for the analysis of foodstuffs [6,22]. e amplicons were produced by conventional end-point PCR as it is a rapid and cheap technique for DNA amplification.…”

“…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)). 6 Journal of Food Quality thuringiensis ssp. Kurstaki is inserted in the transgenic Bt crops to give them insect resistance trait.…”

“…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].…”

An Event-Specific Real-Time PCR Method for Measuring Transgenic Lysozyme Goat Content in Trace Samples

“…In order to establish effective detection methods for transgenic animals and their products, methodologies targeting nucleic acids and proteins are typically used, such as conventional polymerase chain reaction (PCR), multiplex PCR, real-time PCR, digital PCR, enzyme-linked immunosorbent assay (ELISA), and immunochromatographic strips [ 6 , 7 ]. Methods targeting nucleic acids are primarily used for genetically modified organisms (GMOs).…”

Effect of Amount of DNA on Digital PCR Assessment of Genetically Engineered Canola and Soybean Events

“…Accuracy of genetically engineered (GE) trait identification and quantification in non-processed and processed food samples is affected by factors such as sampling/sub-sampling, DNA extraction, DNA quality/purity, DNA amount, and PCR conditions (Alarcon et al 2019;Demeke and Dobnik 2018). The minimum required performance limit (MRPL), which is the lowest level of GE material for satisfactorily reproducible validation of quantitative methods, has been set at 0.1% (European Commission Regulation No.…”