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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2023, Vol. 43 Issue (7): 53-59    DOI: 10.13523/j.cb.2212011
技术与方法     
基于功能核酸的生物传感器对水体中Hg2+的高灵敏检测*
马莉萍,李云霞,聂莹莹,马生龙,郑礴**()
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
Oligonucleotide-based Biosenser for Highly Sensitive Detection of Hg2+ in Aqueous Solution
Li-ping MA,Yun-xia LI,Ying-ying NIE,Sheng-long MA,Bo ZHENG**()
Institute of Sensor Technology, Gansu Academy of Sciences, Key Laboratory of Sensor and Sensing Technology of Gansu, Lanzhou 730099, China
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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-Hg2+-T碱基错配原理构建生物传感器,实现对水体中Hg2+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Hg2+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Hg2+,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Hg2+检测的线性范围为10~800 nmol/L,线性方程为y=0.005 96x+0.881 44(r2=0.991 47),检出限为0.14 nmol/L;10种干扰离子(Fe2+、Co2+、Cu2+、K+、Mg2+、Ca2+、Ag+、Cl-、 SO 4 2 -、 NO 3 -)的存在不影响对Hg2+的检测;在自来水样和黄河水样Hg2+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Hg2+含量的快速检测领域具有良好的应用前景。
关键词： 功能核酸;  Gelred;  生物传感器;  荧光检测;  Hg2+    
Abstract:

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-Hg2+-T base mismatch principle, a new biosensor was constructed to detect the concentration of Hg2+ in water with high sensitivity and selectivity. Methods: A new Gelred/TRO biosensor was constructed by using thymine rich oligonucleotide (TRO) as the specific Hg2+ recognition probe, and Gelred as the fluorescence indicator probe. The sensor is used to detect Hg2+ in water. The influence of many factors on the detection system was studied. Results: Under the optimized detection conditions, the linear range of Hg2+ detection was 10 ~ 800 nmol/L, which was fitted as follows: y=0.005 96x+0.881 44 (r2=0.991 47). The detection limit was 0.14 nmol/L. The presence of ten interfering ions such as Fe2+, Co2+, Cu2+, K+, Mg2+, Ca2+, Ag+, Cl-, SO 4 2 -, and NO 3 - did not affect the detection of Hg2+. The average recovery of Hg2+ 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 Hg2+ in drinking water.
Key words: Oligonucleotide    Gelred    Biosenser    Fluorescent    Hg2+
收稿日期: 2022-12-05 出版日期: 2023-08-03
ZTFLH:  O657.1  
基金资助: 兰州市城关区科技计划(2020-2-1-1);甘肃省科技计划(22JR5RA774);甘肃省科技计划(22CX8GA103);甘肃省科技计划(22JR11RA211)
通讯作者: **电子信箱:760403046@qq.com   
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马莉萍
李云霞
聂莹莹
马生龙
郑礴

引用本文:

马莉萍, 李云霞, 聂莹莹, 马生龙, 郑礴. 基于功能核酸的生物传感器对水体中Hg2+的高灵敏检测*[J]. 中国生物工程杂志, 2023, 43(7): 53-59.

Li-ping MA, Yun-xia LI, Ying-ying NIE, Sheng-long MA, Bo ZHENG. Oligonucleotide-based Biosenser for Highly Sensitive Detection of Hg2+ in Aqueous Solution. China Biotechnology, 2023, 43(7): 53-59.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2212011    或    https://manu60.magtech.com.cn/biotech/CN/Y2023/V43/I7/53

图1  Gelred/TRO传感器检测Hg2+的原理 Detection principle of Gelred/TRO composite probe for Hg2+
图2  荧光发射光谱表征
图3  TRO浓度与荧光强度变化
图4  pH值对传感器荧光信号的影响
图5  反应时间对传感器荧光信号的影响
图6  Gelred/TRO传感器检测不同浓度Hg2+的荧光光谱
图7  荧光强度变化与Hg2+浓度的线性关系
图8  Gelred/TRO传感器检测Hg2+的特异性
Hg2+加标量/
(nmol·L-1)		 自来水 黄河水
测定值/(nmol·L-1) 回收率/% 测定值/(nmol·L-1) 回收率/%
600 576.65±0.54 96.08 556.34±0.21 92.72
700 670.68±0.09 95.81 658.23±0.01 94.03
800 818.89±0.35 102.35 788.51±0.57 98.56
表1  自来水及黄河水中不同浓度Hg2+的检测结果
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