Citation: ZHAO Qiu-Ling, ZHANG You-Hua, YU Xiao-Yan. Fluorescence Aptasensor for Detection of Mercury Ions in Human Urine Using Exonuclease I as Signal Amplification Label[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(7): 1099-1105. doi: 10.11895/j.issn.0253-3820.160029 shu

Fluorescence Aptasensor for Detection of Mercury Ions in Human Urine Using Exonuclease I as Signal Amplification Label

College of Mining Industry Technology, Liaoning Technical University, Huludao 125105, China

Received Date: 12 January 2016
Revised Date: 24 April 2016

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 21506087)

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A novel fluorescence analysis method for fast detection of mercury ions in human urine was established based on DNA recognition and exonuclease I (Exo Ⅰ) assisted signal conversion strategy. The DNA immobilized on the surfaces of the microplates bound specifically with Hg²⁺ to fold into a stable "hairpin" double-stranded structure, thus avoided the hydrolysis by single-stranded DNA specific Exo Ⅰ. Nucleic acid dye SYBR Green I (SG) could insert into the double-stranded "hairpin" part to produce fluorescence signal emission. Based on the above facts, quantitative detection of mercury ions was achieved. The experimental results showed that, when the concentration of avidin coating solution was 50 mg/L and the concentration of the detection DNA immobilized on the microplate was 75 nmol/L, with 5×SG and 1.2 μL of Exo Ⅰ, the optimal analytical performance could be achieved. A good linear relationship between the fluorescence intensity of the system at 520 nm versus the logarithm of mercury ions concentration in the range of 2-500 nmol/L was obtained under the optimal conditions, and the detection limit was 1.5 nmol/L (3σ). The recoveries of mercury in spiked human urine samples were 96.3%-100.5% with the relative standard derivations (RSDs) of 2.1%-4.6%. The method had good selectivity with simple operation, and could be used for the detection of total mercury content after oxidizing other forms of mercury in urine into mercury ions (Ⅱ) using HNO₃-KMnO₄ oxidation method.
- DNA recognition,
- Exonuclease I,
- Mercury ion (Ⅱ),
- Signal amplification strategy,
- Fluorescence analysis
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