A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles

• Published in: Analytica chimica acta 2016-12, Vol.948, p.90-97
• Main Authors: Li, Yaqi, Sun, Li, Qian, Jing, Wang, Chengke, Liu, Qian, Han, En, Hao, Nan, Zhang, Liuping, Cai, Jianrong, Wang, Kun
• Format: Article
• Language: English
• Subjects: Assaying; Caulimovirus - genetics; Deoxyribonucleic acid; DNA; Energy transfer; Fluorescence; Fluorescence Resonance Energy Transfer; Fretting; Genetic modification; Genetically engineered organisms; Genetically modified organisms; Glycine max - genetics; Glycine max - virology; Graphene; Graphite - chemistry; Homogeneous assay; Hybridization; Labeling; Metal Nanoparticles - chemistry; Microscopy, Electron, Transmission; Nanoparticles; Nitrogen; Nitrogen - chemistry; Nitrogen-doped graphene quantum dots; Plants, Genetically Modified - genetics; Plants, Genetically Modified - virology; Polymerase Chain Reaction; Promoter Regions, Genetic; Quantitative analysis; Quantum Dots; Quenching; Reproducibility of Results; Resonance; Silver; Silver - chemistry; Silver nanoparticles; Spectrum Analysis - methods; Target recognition; Transgenic soybean; Viruses
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2016.10.031

In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1–500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection. [Display omitted] •Both NGQDs and AgNPs were selected as the novel FRET donor-acceptor pairs.•The proposed homogeneous FRET assay was developed for CaMV35S detection.•The resulting method could identify 0.5% containing transgenic soybean sample.•This assay was inexpensive, simple and highly sensitive.