Transgenic Plant Detection Using an AuNPs Based SPR Biosensor.

Bartosz F Grześkowiak,Anna Woźniak,Daniel Lipiński,Marlena Szalata,Karol Tuśnio,Ryszard Słomski,Stefan Jurga

doi:10.3390/bios9040116

Bartosz F Grześkowiak, Anna Woźniak + Show 5 more

Open Access

https://doi.org/10.3390/bios9040116

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Abstract

The intensive development and commercialization of genetically modified plants observed over the last decade has led to the development of transgenic detection methods that are rapid and sensitive. Among the strategies used for the detection/monitoring of genetically modified organisms (GMOs), surface plasmon resonance (SPR) meets the necessary criteria. This optical technique measures the changes in the refractive index in the vicinity of thin metal layers (i.e., gold) in response to biomolecular interactions occurring at a flat metal‒solution interface. Additionally, it allows the application of functionalized gold nanoparticles (AuNPs) in SPR research to enhance the signal intensity. In the present study, an SPR method, enhanced by the application of AuNPs, was developed to detect transgenic tobacco plants carrying a Streptococcus mutans antigen. The basis for the detection of the target DNA was the hybridization between the genomic DNA isolated from the leaves, stems, and roots of the transgenic tobacco and the biotinylated oligonucleotide probes immobilized onto a streptavidin (SA) sensor chip. SA-functionalized AuNPs coated with a second type of biotinylated probe were applied to increase the sensitivity of the detection method. Analysis of the results indicated that the constructed SPR-based sensor chip can potentially recognize complementary standard fragments (nonamplified genomic DNA) at concentrations as low as 1 pM. Thus, nonamplified transgenic DNA was detected using a label-free and real-time AuNPs-enhanced SPR biosensing method. This unique approach could be used to detect GMOs with high efficiency, even at a low detection limit, high repeatability, and with less time and a lower cost needed for each analysis.

Highlights

Genetic modification refers to the introduction of an exogenous gene that can express a protein with a specific function
The nucleotide sequences of the biotinylated oligonucleotides
Genomic DNA samples were isolated from the leaves, stems, and roots of transgenic and non-transgenic tobacco

Summary

Introduction

Genetic modification refers to the introduction of an exogenous gene that can express a protein with a specific function. Modified organisms (GMOs) exhibit new traits that can be used for a plethora of applications (e.g., biomedicine, agro-food science, and industry). The development and commercialization of GMOs and their related products will lead to their increased exposure to humans and the environment, and the identification of GMOs may require greater attention. There are two common types of strategies used for GMO detection. The first strategy is based on transgene product targeting indirect protein-based techniques and includes Western blotting, enzyme-linked immunosorbent assays, lateral flow strip tests, and mass spectrometry [1,2].

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