Ren Mengli, Zeng Weili, Li Zhenhua, Cao Shiqin, Liu Chunrong, Ouyang Shuxin, Zhang Tierui, Cui Yanfang, Yuan Hong
Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, China.
Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2020 Oct 15;240:118618. doi: 10.1016/j.saa.2020.118618. Epub 2020 Jun 18.
In the study, CoAl-layered double hydroxide (CoAl-LDH) was prepared as a fluorescence quenching agent to detect DNA molecules. Because of its simple preparation for a large scale, excellent surface effect, good biocompatibility and high fluorescence quenching capability, the effective, rapid, and sensitive DNA detection was realized. The fluorescence quenching efficiency of LDH to 5(6)-carboxyfluorescein attached to single stranded DNA (FAM-ssDNA) was as high as 88%, and after FAM-ssDNA hybridized with the complementary DNA oligonucleotide, that to FAM-dsDNA was about 33%. The quenching mechanisms of LDH for ssDNA and dsDNA were discussed. Phosphate exposed of ssDNA played an important role in quenching effect. Compared to dsDNA, more exposed phosphate groups in ssDNA resulted in the stronger electrostatic interaction between ssDNA and LDH, and thus the higher quenching efficiency. Under optimal conditions, the linear equation was y = 38.26 + 3.37x in a linear relationship of 1-50 nM, and the correlation coefficient R corresponded to 0.999, and the limit of detection was calculated to be 0.79 nM (3σ). Cytotoxicity studies have shown that LDH has good biocompatibility. The study provides an effective, sensitive and safe approach for DNA detection and gives an insight for the design of LDH-based biosensing materials.
在该研究中,制备了钴铝层状双氢氧化物(CoAl-LDH)作为荧光猝灭剂来检测DNA分子。由于其大规模制备简单、具有优异的表面效应、良好的生物相容性和高荧光猝灭能力,实现了有效、快速且灵敏的DNA检测。LDH对附着于单链DNA的5(6)-羧基荧光素(FAM-ssDNA)的荧光猝灭效率高达88%,而在FAM-ssDNA与互补DNA寡核苷酸杂交后,其对FAM-双链DNA(FAM-dsDNA)的猝灭效率约为33%。讨论了LDH对单链DNA和双链DNA的猝灭机制。单链DNA暴露的磷酸基团在猝灭效应中起重要作用。与双链DNA相比,单链DNA中更多暴露的磷酸基团导致单链DNA与LDH之间更强的静电相互作用,从而具有更高的猝灭效率。在最佳条件下,线性方程为y = 38.26 + 3.37x,线性范围为1 - 50 nM,相关系数R为0.999,计算得到的检测限为0.79 nM(3σ)。细胞毒性研究表明LDH具有良好的生物相容性。该研究为DNA检测提供了一种有效、灵敏且安全的方法,并为基于LDH的生物传感材料的设计提供了思路。