Biosensor for detecting mercury ions on basis of fluorescence

A biosensor and fluorescence detection technology, applied in the field of biosensors, can solve the problems of long detection period, high cost, low specificity and sensitivity, etc., and achieve the effect of simple preparation method, stable performance and sensitive detection

Inactive Publication Date: 2018-07-27
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the problems of relatively low specificity and sensitivity, high cost and long detection period of the method for detecting mercury ions in the prior art, the present invention provides a G-quadruple-based method with high specificity and sensitivity, low cost and fast detection speed. Body Quenched Silver Cluster Fluorescence to Detect Mercury Ion Biosensor

Method used

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  • Biosensor for detecting mercury ions on basis of fluorescence
  • Biosensor for detecting mercury ions on basis of fluorescence
  • Biosensor for detecting mercury ions on basis of fluorescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of D1-AgNCs.

[0033] Configure PB buffer solution (concentration is 20mM), PB buffer solution is composed of disodium hydrogen phosphate and sodium dihydrogen phosphate, weigh 0.7163g disodium hydrogen phosphate and 0.3120g sodium dihydrogen phosphate respectively, and make 100ml solution each , and then take a part of disodium hydrogen phosphate and mix a part of sodium dihydrogen phosphate, adjust the pH value of the mixed solution to 6.5 and then set aside.

[0034] Formulating AgNO 3 Concentration of 2mM, volume of 1mL, AgNO 3 Easy to decompose when exposed to light, ready-to-use and ready-to-use, configure AgNO 3 Wrap the centrifuge tube with tinfoil first.

[0035] Take a 1 mL centrifuge tube, add 76 μL of PB (20 mM), add 15 μL of D1 (final concentration is 15 μM), add 4.5 μL of AgNO 3 (2mM), shake for 1min, and put in a refrigerator at 4°C for 30min. Prepare NaHBO during 4 Concentration 2mM, volume 1mL, NaHBO 4 Ready-to-use and ready...

Embodiment 2

[0037] Example 2 Effects of different concentrations D1 on the detection of mercury ions.

[0038] (1) Dilute the D1-AgNCs solution in Example 1 to a final concentration of 0.4 μM, 0.6 μM, 0.8 μM, 1.0 μM, 1.2 μM, and 1.4 μM, and then measure the fluorescence intensity respectively. Take 2 μL and add them to 7 centrifuges tube, then add 2 μL of 15 μM D2 chain to each tube, 8 μL of 5×PBS, 2 μL of heme (1 μM), and finally 2 μL of Hg 2+ solution (3μM), shaken for 30s, and placed in a water bath at 37°C for 1h.

[0039] (2) After 1 h, take out the mixed solution from the water bath, and then measure its fluorescence intensity.

[0040] Take the concentration of D1 as the abscissa, and the fluorescence intensity as the ordinate, as figure 2 . It can be seen from the figure that the detected fluorescence signal intensity decreases as the concentration of D1 increases in the range of 0.4-1.4 μM, and when the concentration exceeds 1.0 μM, the fluorescence intensity tends to be stab...

Embodiment 3

[0041] Example 3 Effects of different concentrations of D2 on the detection of mercury ions.

[0042] (1) To measure the fluorescence intensity of D1-AgNCs sample S4 in Example 1, take 2 μL of S4 with a concentration of 15 μM and add them to 7 centrifuge tubes, and then add 2 μL of D2 chains with different concentrations (final concentrations are 0.4 μM, 0.6 μM, respectively). μM, 0.8μM, 1.0μM, 1.2μM, 1.4μM), add 8μL of 5×PBS to each tube, 2μL of heme (1μM), and finally 2μL of Hg 2+ The solution (3μM) was added to the centrifuge tube, shaken for 30s, and placed in a water bath at 37°C for 1h.

[0043] (2) Take out the mixed solution from the water bath after 1 h, and then measure its fluorescence intensity.

[0044]Take the D2 chain concentration as the abscissa, and the fluorescence intensity as the ordinate, as image 3 . It can be seen from the figure that the detected fluorescence signal intensity decreases as the concentration of D2 chain increases in the range of 0.4-...

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Abstract

The invention provides a biosensor for detecting mercury ions on the basis of fluorescence. The biosensor comprises components as follows: a DNA-containing silver cluster, DNA capable of forming a G-quadruplet, potassium ions and heme. The biosensor is mild in detection reaction conditions, the reaction speed is increased, the operation complexity is reduced, and rapid, simple and sensitive detection of a target object is realized. The preparation method of the biosensor is simple, the performance is stable, the repeatability of the silver cluster is good, and the biosensor is mainly applied to detection of food safety and mercury ions in water and actual application of biosensor industrialization. A process for manufacturing the biosensor is low in cost and applicable to the cheap requirement in industrialization.

Description

technical field [0001] The invention relates to a biosensor for fluorescence detection of mercury ions, belonging to the technical field of biosensors. Background technique [0002] Metallic mercury poisoning is often caused in the form of mercury vapor. Because mercury vapor is highly diffusible and fat-soluble, it enters the alveoli through the respiratory tract and is transported to the whole body through blood circulation. After the metal mercury in the blood enters the brain tissue, it is oxidized into mercury ions, gradually accumulates in the brain tissue, and when it reaches a certain amount, it will cause damage to the brain tissue. Another part of the mercury ions is transferred to the kidneys. Therefore, the clinical manifestations of chronic mercury poisoning are mainly neurological symptoms, such as headache, dizziness, numbness and pain in limbs, muscle tremor, and ataxia. Excitement is a special mental state of chronic mercury poisoning, manifested as irrit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/643
Inventor 王玉张雪刘素黄加栋唐丹
Owner UNIV OF JINAN
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