基于T碱基错配非标记荧光法检测Hg2+的新方法  

A new method for detecting Hg2+ with T-base mismatched label-free fluorescence method

作　　者：陆文涛 王蓉 侯少龙 张红鸽 LU Wen-tao;WANG Rong;HOU Shao-long;ZHANG Hong-ge(College of Chemistry and Chemical Engineering,Baoji University of Arts and Sciences,Baoji 721013,Shaanxi,China)

机构地区：[1]宝鸡文理学院化学化工学院,陕西宝鸡721013

出　　处：《宝鸡文理学院学报（自然科学版）》2020年第2期25-31,共7页Journal of Baoji University of Arts and Sciences(Natural Science Edition)

基　　金：陕西省教育厅重点实验室项目(18JS007);宝鸡文理学院重点实验室项目(ZK2018060)。

摘　　要：目的建立一种高选择性的、非标记、操作简单的检测Hg2+的荧光分析方法。方法利用紫外可见吸收光谱法,在260 nm波长下测吸光度,对DNA进行定量分析,准确测定实验所用DNA片段的浓度;利用荧光光谱法,在激发波长为445 nm,扫描范围为455~650 nm下,检测单碱基T/T错配的双链DNA探针加入核黄素和Hg2+前后,核黄素的荧光信号的变化。结果单碱基错配的双链DNA能有效检测10~200 nM Hg2+。该体系的荧光比率(F/F0)与Hg2+浓度具有良好的线性关系,其线性方程为F/F0=0.0009C(nmol/L)+0.0049,相关系数r2=0.9930,检出限为3.3 nM(S/N=3)。相对标准偏差(RSD)小于1.37%;样品回收率为100.8%~103.3%。结论该方法可用于环境水样中Hg2+的测定。Purposes—To establish a highly selective,label-free,simple-operation fluorescence detection method for detecting Hg2+.Methods—The absorbance was measured with ultraviolet-visible absorption spectroscopy at a wavelength of 260 nm for quantitative analysis of DNA and accurate determination of the concentration of the DNA fragments used in the experiment.Fluorescence spectroscopy was used to detect the changes in fluorescence signal of riboflavin at excitation wavelength of 445 nm and scanning range of 455~650 nm before and after the addition of riboflavin and Hg2+to single-base T/T mismatched double-stranded DNA probes.Result—Single-base mismatched double stranded DNA can effectively detect 10~200 nM Hg2+.The fluorescence ratio(F/F0)of the system has a good linear relationship with the Hg2+concentration.The linear equation is F/F0=0.0009C(nmol/L)+0.0049,the correlation coefficient r2 was equal to 0.9930 and the detection limit was 3.3 nM(S/N=3).The relative standard deviation(RSD)was less than 1.37%,and the recovery ranged from 100.8%to 103.3%.Conclusion—This method can be used for the determination of Hg2+in water samples in bad environment.

关 键 词：非标记法 T/T碱基错配 荧光分析法 Hg2+ 

分 类 号：O661.1[理学—分析化学] S951.42[理学—化学]