Citation: LEI Zhao-Jing, ZHANG Cun-Zheng, HU Qiu-Hui, LIU Yuan, ZHANG Qiang, LIU Xian-Jin. Aptamer-Based Fluorescence Assay for Hg²⁺ Determination[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(12): 1827-1831. doi: 10.3724/SP.J.1096.2012.20127 shu

Aptamer-Based Fluorescence Assay for Hg²⁺ Determination

1.
¹ College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China;

Corresponding author: LIU Xian-Jin,
Received Date: 15 March 2012
Available Online: 18 June 2012
Fund Project: 本文系江苏省农业科技自主创新项目(No.CX(10)236) (No.CX(10)236)苏州市科技支撑项目(No.SN201129) (No.SN201129)中国博士后基金(No.20090451177) (No.20090451177)江苏省博士后科研资助计划(No.0802023B)资助项目 (No.0802023B)

A RNA aptamer modified to DNA (N1) that could bind to the target of Ni²⁺ was found that could bind specifically to Hg²⁺ ion with typical secondary structure. Thereafter, several DNA aptamers with a certain conformation of sequence mutation and splice were designed in order to increase the property of Hg²⁺ binding, and Aptamer-based fluorescence assay for the Hg²⁺ ion determination was developed. The addition of the Hg²⁺ to a mixture containing the duplex of a fluorophore labeled aptamer and a quencher-modified antisense nucleic acid (Q2) would force the release of Q2 from labeled aptamer, which was spontaneously accompanied by the increase of fluorescence intensity and made Hg²⁺ in quantitative analysis. It was shown that stable labeled aptamer DNA and Q2 hydrogen bonds formed at concentration ration of 1:3(labeled aptamer and Q2, respectively), under the condition of 5 min denaturation in 94℃ and 30 min renaturation in room temperature (25℃) in the fluorescence assay preparation. For aptamer N1, the fluorescence assay results showed that linear response toward Hg²⁺ concentration with fluorescence quenching system ranged from 1.25 mg/L to 20 mg/L, with the limit of detection 0.625 mg/L. In order to improve the sensitivity of aptamer N1, 4 oligonucleotides (N4, N5, N6, N7) were designed and synthesized based on the structure of N1. The results showed that N5 had the best affinity and specificity to Hg²⁺ which displayed the linear range for the Hg²⁺ concentration detection was 0.156 mg/L to 2.5 mg/L with a detection limit of 78 μg/L.
- Aptamer,
- Fluorescence,
- Mercury(Ⅱ),
- Detection
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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

Aptamer-Based Fluorescence Assay for Hg2+ Determination

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通讯作者: 陈斌, bchen63@163.com

Aptamer-Based Fluorescence Assay for Hg²⁺ Determination