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Mercury ion fluorescence detection method and kit

A fluorescence detection, mercury ion technology, applied in the direction of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of poor selectivity, poor water solubility of the sensor, slow response speed, etc., and achieve the effect of low cost, short sequence and small response

Inactive Publication Date: 2013-09-11
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this type of sensor has poor water solubility, Hg 2+ The disadvantages of slow response and poor selectivity

Method used

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  • Mercury ion fluorescence detection method and kit
  • Mercury ion fluorescence detection method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Fluorescence-labeled and non-fluorescent-labeled MSO dosage optimization design

[0034]Take 2.5 μl of different concentrations (10nM, 100nM, 1μM, 10μM) of fluorescein (FAM)-labeled and non-fluorescent-labeled MSO, add 265μl of 20mM Tris-HCl buffer, react at room temperature for 5 minutes, and use RF-5301PC fluorescence spectrometry The photometer scans the fluorescence wavelength curve. Fluorescence spectrum measurement conditions of the fluorescence spectrophotometer: xenon lamp excitation, slit width 10nm, response time Auto. The excitation wavelength is 496nm, and the scanning range of the emission spectrum is 515-600nm. The measurement is performed at room temperature with a 350μL microcuvette, and the fluorescence value at 522nm is recorded. The fluorescence values ​​at 522nm of different concentrations of fluorescently labeled MSO were recorded for comparison. The result is as figure 1 : When the concentration of fluorescently labeled and non-fluores...

Embodiment 2

[0035] Embodiment 2 buffer optimization design

[0036] Prepare Tris-HCl buffer containing metal ions and Tris-HCl buffer without metal ions. After taking 2.5 μl of 1 μM fluorescently-labeled and non-fluorescently-labeled MSO, these two buffers were added to scan the fluorescence wavelength curve, and the fluorescence value at a wavelength of 522 nm was recorded. Finally, 5 μl of 0.5 g / L GO was added, reacted at room temperature for 5 minutes, and then scanned the fluorescence wavelength to record the change of fluorescence value. figure 2 Among them, when the Tris-HCl buffer containing metal ions was added to the reaction system, the fluorescence quenching efficiency reached (55.191±5.4)% on average before and after adding GO, which was relatively stable; when the Tris-HCl buffer containing no metal ions solution was added to the reaction system, before and after adding GO, the fluorescence quenching efficiency reached (10.108±2.2)% on average, which was much lower than the...

Embodiment 3

[0037] Embodiment 3GO and Hg 2+ Exploring the sequence of adding samples

[0038] After taking 2.5 μl of 1 μM fluorescently labeled and unfluorescently labeled MSO, first add 5 μl of 0.5 g / L GO, 265 μl of 20 mM Tris-HCl buffer, and then add Hg 2+ and add Hg first 2+ , 265μl of 20mM Tris-HCl buffer solution, and then add 5μl of 0.5g / LGO in these two ways for time kinetic detection, continuous detection for 15 minutes, and observe the change of fluorescence intensity. At the same time, each group uses water group as the control group. The optimal sequence of adding samples for the detection of mercury ions by fluorescence method. from image 3 The results showed that adding GO first and then adding Hg 2+ The signal-to-noise ratio of the group increases with the prolongation of time, and the S / N at 15 minutes can reach 1.476±0.123, indicating that after adding mercury ions, the fluorescence does recover, and the experimental results are better than adding Hg 2+ Add GO group ...

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Abstract

The invention relates to a mercury ion fluorescence detection method and a kit for rapid detection of mercury ions in water. Specifically, original MSO (mercury-specific oligonucleotide) is optimized and modified, fluorescence modified and non-fluorescence modified optimized MSO is adsorbed to the surface of GO (graphene oxide) by means of a non covalent bond's (pi-pi stacking) acting force, and MSO and mercury ions are utilized to form a T-Hg<2+>-T structure, so that fluorescently-labeled MSO can be released from the GO surface, and fluorescence signals can be restored. By detecting the change of fluorescence signals in a solution, quantitative detection of mercury ions in the solution can be realized. The method provided in the invention can realize convenient analysis of mercury ions without requiring any special condition. The lowest detection limit is 10pM, the sensitivity of original methods is enhanced by 18 times, and the the cost of DNA synthesis is reduced. Thus, the invention provides a simple, efficient, sensitive and cost-effective solution for monitoring mercury ion pollution in water, and the solution is easy to promote and use.

Description

technical field [0001] The invention belongs to the technical field of environmental detection, and relates to a mercury ion fluorescence detection method based on mercury ion-specific oligonucleotide probe (mercury-specific oligonucleotide, MSO) and graphene oxide (Graphene oxide, GO) and its application in water A kit for the rapid detection of mercury ions. Background technique [0002] In recent years, with the rapid development of modern industry and agriculture, environmental pollution and poisoning incidents caused by mercury broke out frequently. Among them, the problem of mercury pollution in water bodies in my country is relatively prominent. According to the survey by the Ministry of Agriculture, the area of ​​sewage irrigation areas in my country is about 140×10 4 hm 2 , the area of ​​land polluted by heavy metals accounted for 64.8% of the total area of ​​pollution, among which the area of ​​pollution such as mercury was the largest. There are currently about...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 孙涛何珏张林志谢晏讷项序武洪小兰姚小燕吴文鹤吕建新
Owner WENZHOU MEDICAL UNIV
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