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Establishment and Application of Multiplex Polymerase Chain Reaction for Simultaneous Detection of Transgenic Potatoes
The results of specificity test of primers among events and species. The samples of 10% and 0% genetically modified (GM) potato (EH92-527-1, AM04-1020, 2-1), soybean, and maize were used and four target sequences including EH92 (450 bp), AM04 (238 bp), 2-1 (357 bp), and UDP-glucose pyrophosphorylase (UGPase) (161 bp) were detected by multiplex polymerase chain reaction (mPCR) simultaneously.
The results of specificity test of primers among events and species. The samples of 10% and 0% genetically modified (GM) potato (EH92-527-1, AM04-1020, 2-1), soybean, and maize were used and four target sequences including EH92 (450 bp), AM04 (238 bp), 2-1 (357 bp), and UDP-glucose pyrophosphorylase (UGPase) (161 bp) were detected by multiplex polymerase chain reaction (mPCR) simultaneously.

Author:Yuan-Kai Tu, Yu-Wei Feng, Lit-Fu Chan, Shu Chen, Yen-Chun Lin, and Han-Wei Chen*

Abstract:

    Potato (Solanum tuberosum L.) is known as one of the most important food crops in the world and has high application in industry and feed. In this study, we present a multiplex polymerase chain reaction (mPCR) method for simultaneous detection of genetically modified (GM) potatoes. Quadruplex PCR assay targeting three event-specific sequences and the taxon-specific endogenous reference gene (UDP-glucose pyrophosphorylase; UGPase) were established. All potato DNA samples generated the expected PCR products, and the detection method performed robustly and specifically with various combinations of mixed GM potato events. The limit of detection (LOD) of each single GM sample in our mPCR detection assay was 0.5%. Furthermore, a proficiency test returned from four research sectors also indicated that our detection method had good reproducibility and repeatability. We also developed a simple, rapid and cost-efficient nucleic acid extraction method coupled with multiplex PCR and followed capillary electrophoresis. These results suggest that our methods are suitable for detecting multiplex PCR targets and have the potential for screening unknown genetically modified potato samples through a simple procedure.

Key words:Potato, mPCR, Detecting method

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Journal of
Taiwan Agricultural Research
Vol. 73, No.1 (2024) ISSN: 2790-086X Vol. 73, No.1 (2024)
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