A magnetic-separation-dual-targets fluorescent biosensor was fabricated to detect terminator nopaline synthase (TNOS) and promoter of cauliflower mosaic virus 35s (P35S) in transgenic soybean based on incorporation of bicolor CdTe quantum dots carried by silica nanospheres. In this protocol, the fixed probes for TNOS or P35S were magnetized firstly with Fe3O4@Au magnetic nanosphere by Au-S covalent bonding to achieve magnetized probes. Meanwhile, the capture probes for TNOS or P35S were functionalized with green or red fluorescent microspheres respectively to obtain fluorescently-labeled probes, which could emit relative strong green or red fluorescent signal. Two terminals of TNOS or P35S were recognized by magnetized probes and fluorescently-labeled probes respectively to form the sandwiched structures in the process of biosensor development subsequently, and it was separated by a magnet instantly. The fluorescence intensities of remnant supernatant were measured and analyzed accordingly to achieve simultaneous detection of TNOS and P35S. This biosensor exhibited a good dynamic range, low limit of detection and excellent selectivity in detecting transgenic soybean.
Keywords: Dual targets; Fluorescent quantum dots; Genetically modified organism; Magnetic nanosphere.
Copyright © 2020 Elsevier B.V. All rights reserved.