基于功能核酸的生物传感器对水体中Hg^(2+)的高灵敏检测  

Oligonucleotide-based Biosenser for Highly Sensitive Detection of Hg^(2+) in Aqueous Solution

作　　者：马莉萍[1] 李云霞[1] 聂莹莹[1] 马生龙[1] 郑礴[1] MA Li-ping;LI Yun-xia;NIE Ying-ying;MA Sheng-long;ZHENG Bo(Institute of Sensor Technology,Gansu Academy of Sciences,Key Laboratory of Sensor and Sensing Technology of Gansu,Lanzhou 730099,China)

机构地区：[1]甘肃省科学院传感技术研究所甘肃省传感器与传感技术重点实验室,兰州730099

出　　处：《中国生物工程杂志》2023年第7期53-59,共7页China Biotechnology

基　　金：兰州市城关区科技计划(2020-2-1-1);甘肃省科技计划(22JR5RA774、22CX8GA103、22JR11RA211)资助项目。

摘　　要：目的:汞是一种广泛存在于环境中的剧毒污染物,在较低浓度就能对人体造成严重危害;基于T-Hg^(2+)-T碱基错配原理构建生物传感器,实现对水体中Hg^(2+)浓度的高灵敏、高选择性检测。方法:筛选富含胸腺嘧啶的寡核苷酸(thymine rich oligonucleotide,TRO)作为Hg^(2+)特异识别探针,以核酸荧光染料Gelred作为荧光指示探针,构建Gelred/TRO生物传感器检测水体中Hg^(2+),并分析多种因素对检测体系的影响。结果:优化检测条件下,Gelred/TRO传感器对Hg^(2+)检测的线性范围为10~800 nmol/L,线性方程为y=0.00596x+0.88144(r2=0.99147),检出限为0.14 nmol/L;10种干扰离子(Fe^(2+)、Co^(2+)、Cu^(2+)、K^(+)、Mg^(2+)、Ca^(2+)、Ag+、Cl^(-)、SO_(4)^(2-)、NO3-)的存在不影响对Hg^(2+)的检测;在自来水样和黄河水样Hg^(2+)加标检测实验中,平均回收率分别为98.08%和95.1%。结论:基于功能核酸的生物传感器操作简便、灵敏度高、特异性好、检测限低,在饮用水Hg^(2+)含量的快速检测领域具有良好的应用前景。Objective:Mercury is a highly toxic and widespread pollutant in the environment,which can cause harm to human body even in low concentration.Based on the T-Hg^(2+)-T base mismatch principle,a new biosensor was constructed to detect the concentration of Hg^(2+)in water with high sensitivity and selectivity.Methods:A new Gelred/TRO biosensor was constructed by using thymine rich oligonucleotide(TRO)as the specific Hg^(2+)recognition probe,and Gelred as the fluorescence indicator probe.The sensor is used to detect Hg^(2+)in water.The influence of many factors on the detection system was studied.Results:Under the optimized detection conditions,the linear range of Hg^(2+)detection was 10~800 nmol/L,which was fitted as follows:y=0.00596x+0.88144(r2=0.99147).The detection limit was 0.14 nmol/L.The presence of ten interfering ions such as Fe^(2+),Co^(2+),Cu^(2+),K^(+),Mg^(2+),Ca^(2+),Ag+,Cl^(-),SO_(4)^(2-),and NO3-did not affect the detection of Hg^(2+).The average recovery of Hg^(2+)in tap water samples and the Yellow River were 98.08%and 95.1%,respectively.Conclusion:This method has the advantages of simple operation,high sensitivity,good specificity and low detection limit.It has a good application prospect in the rapid determination of Hg^(2+)in drinking water.

关 键 词：功能核酸 Gelred 生物传感器 荧光检测 Hg^(2+) 

分 类 号：O657.1[理学—分析化学]