Functional Nucleic Acid Based Fluorescent Biosensing Method for Hg2+ Detection in Water Samples
ZHU Xi-yu1, 2, WANG Ruo-yu2, ZHOU Xiao-hong2*, TAN Ai-juan1*, WEN Xiao-gang3*
1. College of Life Sciences, Guizhou University, Guiyang 550025, China
2. State Key Joint Laboratory of Environmental Simulation and Pollutants Control, School of Environment, Tsinghua University, Beijing 100084, China
3. Nanhu College, Jiaxing University, Jiaxing 314001, China
摘要： 二价汞离子作为一种重要的环境污染物，一直以来受到国内外广泛关注。基于T-T错配的Hg2+检测极大依赖于汞选择性寡聚核苷酸（MSO）的设计，利用SYBR Green I对目前所报道的汞离子探针进行了优化，研究了若干种MSO探针与Hg2+的结合响应，在对探针二级结构进行分析和讨论的基础上，提出最优的富T探针序列，并由此建立了一种基于SYBR Green I的水中汞离子快速、便捷的荧光检测方法。最终测得3种实际水样的加标回收率在82.8％~101.8％之间，相对标准偏差小于15%，表明该方法受环境基质的影响较小，可应用于实际水样中的汞离子检测。
关键词：二价汞离子；T-Hg2+-T复合物；SYBR Green I；荧光分析法
Abstract：As an important environmental contaminant, mercury(Ⅱ) has caused worldwide concern. The T-T mismatch based detection for Hg2+ greatly depends on the design of mercury specific oligonucleotide (MSO). This paper optimized the currently reported MSOs by using SYBR Green I. Based on the binding response analysis between several MSOs and Hg2+and further discussion on the secondary structure of sequences, the optimum T-rich sequences was proposed. Hence a fluorescent detection method towards Hg2+ in water samples based on SYBR Green Ⅰ was established. Consequently, the recovery ratios in spiked real water samples ranged from 82.8% to 101.8% with the relative standard derivation of less than 15%. Results show that this method was slightly affected by the environmental matrix, providing accurate detection of mercury ions in real water samples.
[1] Monitoring A. Available on http://www.amap.no/documents/doc/technical-background-report-for-the-global-mercury-assessment-2013/848 (accessed on 17 June 2014): 263.
[2] Lollar B S. Environmental Geochemistry: Elsevier, 2005.
[3] Ono A, Togashi H. Angewandte Chemie International Edition, 2004, 43(33): 4300.
[4] Jayasena S D. Clinical Chemistry, 1999, 45(9): 1628.
[5] Xing Y-P, Liu C, Zhou X-H, et al. Scientific Reports, 2015, 5: 8125.
[6] Wang R, Zhou X, Shi H. Biosens Bioelectron, 2015, 74: 78.
[7] Zhang J R, Huang W T, Zeng A L, et al. Biosensors and Bioelectronics, 2015, 64(Supplement C): 597.
[8] Qiu H, Wu N, Zheng Y, et al. International Journal of Nanomedicine, 2015, 10: 147.
[9] Xu H, Zhu X, Ye H, et al. Chemical Communications, 2011, 47(44): 12158.
[10] Teh H B, Wu H, Zuo X, et al. Sensors and Actuators B: Chemical, 2014, 195: 623.
[11] Wang R, Zhou X, Shi H, et al. Biosens Bioelectron, 2016, 78: 418.
[12] Wang J, Liu B. Chemical Communications, 2008，(39): 4759.
[13] Zipper H, Brunner H, Bernhagen J, et al. Nucleic Acids Research, 2004, 32(12): e103.
[14] Zhou Y, Deng M, Du Y, et al. Analyst, 2011, 136(5): 955.
[15] Li D, Wang J, Zheng G, et al. Nanotechnology, 2013, 24(23): 235502.
[16] Niu S-Y, Li Q-Y, Ren R, et al. Analytical Letters, 2010, 43(15): 2432.
[17] Xu L, Yin H, Ma W, et al. Biosensors and Bioelectronics, 2015, 67(Supplement C): 472.
[18] Zhu Z, Xu L, Zhou X, et al. Chemical Communications, 2011, 47(28): 8010.
