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Simultaneous detection of TNOS and P35S in transgenic soybean based on magnetic bicolor fluorescent probes.
Talanta ( IF 6.1 ) Pub Date : 2020-01-20 , DOI: 10.1016/j.talanta.2020.120764
Yaqi Li 1 , Nan Hao 2 , Shilong Luo 3 , Qian Liu 2 , Li Sun 4 , Jing Qian 2 , Jianrong Cai 4 , Kun Wang 5
Affiliation  

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.

中文翻译:

基于磁性双色荧光探针同时检测转基因大豆中的TNOS和P35S。

制备了磁分离双靶荧光生物传感器,通过掺入二氧化硅纳米球携带的双色CdTe量子点,检测转基因大豆中终止子胭脂碱合酶(TNOS)和花椰菜花叶病毒35s(P35S)的启动子。在该协议中,首先通过Au-S共价键将Fe3O4 @ Au磁性纳米球磁化TNOS或P35S的固定探针,以实现磁化探针。同时,分别用绿色或红色荧光微球对TNOS或P35S的捕获探针进行功能化,以获得可以发出相对较强的绿色或红色荧光信号的荧光标记探针。TNOS或P35S的两个末端分别被磁化探针和荧光标记的探针识别,在随后的生物传感器开发过程中形成了夹层结构,并立即被磁体分离。测量并分析残余上清液的荧光强度,以同时检测TNOS和P35S。该生物传感器表现出良好的动态范围,低检测限和在检测转基因大豆中的优异选择性。
更新日期:2020-01-21
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