GMO analysis technique based on PCR and real-time PCR methods

Abstract

The use of genetically modified organisms (GMOs) is promising for overcoming the shortage of food products in the world and solving the problem of hunger arising in various regions. At the same time, the use of GMOs has become a cause of debate, as the safety of consuming GMO products for human health remains unproven. The risks associated with GMOs cause public concern, which has led to the restriction of the use of GMOs and their products in many countries and the need for constant control of their content in food products. This study describes methods based on the polymerase chain reaction (PCR), such as Real-time PCR and PCR with electrophoretic separation of amplificates, which are generally accepted in GMO analysis. In order to control the presence of GMOs in food raw materials, feed and finished products of plant and animal origin, screening is carried out for the presence of the most common genetic engineering structures used during the creation of GMOs: CaMV 35S promoter and NOS terminator. Both Real-time PCR and PCR with electrophoretic separation of amplificates allow to establish the presence of GMOs with high accuracy, and Real-time PCR is also used to determine the concentration of GMOs in the studied samples. The work presents a typical electrophoresis with visualization of the obtained PCR fragments that required electrophoretic separation and fragments that were synthesized in the Real-time PCR reaction, and determined the approximate sizes of the obtained fragments relative to the pBR322 DNA-MspI molecular weight marker. The approach described in this study, based on the use of PCR techniques, can be successfully used for GMO analysis of all groups of raw materials and finished products of animal and plant origin, and is also well adapted for the detection of various genetic engineering structures, not limited to the CaMV 35S promoter and terminator NOS, which makes it possible to increase the efficiency of such analysis and continue its application in relation to new genetic engineering structures used during the creation of GMOs. Keywords: genetically modified organisms (GMOs), PCR methods, GMO analysis, raw materials and finished products of plant and animal origin