Development of Gene-Specific Real-Time PCR Screening Method for Detection ofcp4-epspsGene in GM Crops

Abstract

Among the genetically modified (GM) crops that are being approved for commercialization, herbicide resistant crops, especially those harboring cp4-epsps, have a considerable contribution. Gene-specific methods can be used to screen the presence of GMOs. To establish an effective qualitative and quantitative screening method, a set of primers were designed considering the cp4-epsps sequence. The specificity, the limit of detection, the efficiency, and the ability to quantify the GMO content were tested in GM cotton, soybean, and canola events. The results demonstrated that the primers can specifically detect cp4-epsps GM crops. The limit of detection was found to be 0.4 ng /μl DNA per PCR reaction with the ability to detect 1-16 copies of the haploid genome of each GM event. The efficiency of this screening method (which was 94-110 % with an R2 higher than 0.96) indicated that these new primers can be applied to the screening of GM samples that contain the cp4-epsps gene. Also, the gene-specific real-time PCR screening method could be successfully developed for qualification of different types of GM cotton, soybean and canola events with the construction of a serial dilution ranging from 10 % to 1 %.

A dual-RPA based lateral flow strip for sensitive, on-site detection of CP4- EPSPS and Cry1Ab/ Ac genes in genetically modified crops

“…To fully evaluate the program constructed in this study, common transgenic assays and their detection capabilities were listed in Table 1. By comparison, overall, conventional nucleic acid-based assays [24,25] tend to be more sensitive than protein-based methods [26][27][28][29]. Nucleic acid amplification is the core of most nucleic acid-based methods.…”