Development and verification of new reference plasmid (pUC_GM-SB) for the DNA-based detection of genetically modified sugar beet H7-1

Food Sci Biotechnol. 2024 May 1;33(14):3291-3297. doi: 10.1007/s10068-024-01572-6. eCollection 2024 Nov.

Abstract

The sugar beet is the second-largest sugar-producing crop. Genetically modified (GM) sugar beet, which have herbicide-resistant, have been developed to increase production and comprise over 90% of the market share. This study describes qualitative and quantitative PCR methods for the GM sugar beet H7-1 with reference plasmid (pUC_GM-SB) containing an endogenous gene (GS2) and an event-specific gene for H7-1 that served as a positive control for PCR. The detection limit of qualitative PCR was approximately 10 copies of the reference plasmid and 0.05% in spiked samples. In the case of quantitative PCR, the detection limit was five copies of the reference plasmid. Regarding repeatability, the standard deviation and relative standard deviation were found to range from 0.11 to 0.24 and from 0.23% to 0.99%, respectively. This study provides food safety assurance for imported GM sugar beet H7-1 using the reference plasmid and supports efficient detection methods.

Supplementary information: The online version contains supplementary material available at 10.1007/s10068-024-01572-6.

Keywords: Event-specific primer; GM sugar beet; Qualitative PCR; Quantitative PCR; Reference plasmid.