Colorimetric biosensor for detecting mercury ions and preparation method thereof

A biosensor and mercury ion technology, applied in the field of biosensors, can solve the problems of low specificity and sensitivity, high cost, long detection cycle, etc., and achieve the effects of improved sensitivity, low detection limit and fast detection speed

Inactive Publication Date: 2017-06-20
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve 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 chain-based method with high specificity and sensitivity, low cost and fast detection speed. A colorimetric biosensor for displacement and detection of mercury ions by gold nanoparticles

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  • Colorimetric biosensor for detecting mercury ions and preparation method thereof
  • Colorimetric biosensor for detecting mercury ions and preparation method thereof
  • Colorimetric biosensor for detecting mercury ions and preparation method thereof

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preparation example Construction

[0046] The preparation method of described biosensor comprises the following steps:

[0047] (1) Preparation of gold nanoparticles;

[0048] (2) Modify Probe4 to the surface of gold nanoparticles;

[0049] (3) Mix the labeled gold nanoparticles solution with the homogeneous reaction solution.

[0050]The preparation method, preferably the preparation steps of gold nanoparticles are as follows:

[0051] ① Install the required instruments and add 200ml of ultrapure water to the three-necked flask (be careful not to let dust fall into the three-necked flask);

[0052] ② Take 500uL (0.04g / ml) HAuCl 4 In a single packaged centrifuge tube, use a pipette gun to take 500ul and 200ml of ultrapure water, stir and heat at a stirring speed of about 450 rpm until boiling;

[0053] ③Under stirring conditions, take 3ml of 1% trisodium citrate solution and quickly add it into the solution. Within a few minutes, the color of the solution changes from light yellow to wine red. After continu...

Embodiment 1

[0064] The steps for modifying Probe4 to the surface of gold nanoparticles are as follows:

[0065] a. Take 1 mL nano-gold solution in a centrifuge tube, centrifuge for 10 min, and centrifuge the two tubes at the same time for later use. Centrifuge until the supernatant is colorless and transparent, remove the supernatant, and add 300 μL of sterile water to concentrate the nano-gold solution to 3 nM. Transfer to a 1 mL glass bottle and seal with tinfoil;

[0066] b. After standing at room temperature for 30 minutes, add 150 μL of -SH-modified substrate probe (Probe4) with a concentration of 30 μM, mix well, and place at 4 °C for 24 hours;

[0067] c. Slowly add 50 μL of PBS buffer several times, add magnets (soaked in aqua regia the day before) and stir for 10 min, then continue to add 27 μL of PBS buffer. Take out the magnet and place it at 4°C for 48 h;

[0068] d. Transfer the labeled gold nanoparticles solution to a centrifuge tube, add sterilized water to 1 mL, centrif...

Embodiment 2

[0078] The steps for modifying Probe4 to the surface of gold nanoparticles are as follows:

[0079] a. Take 1 mL nano-gold solution in a centrifuge tube, centrifuge for 10 min, and centrifuge the two tubes at the same time for later use. Centrifuge until the supernatant is colorless and transparent, remove the supernatant, and add 300 μL of sterile water to concentrate the nano-gold solution to 3 nM. Transfer to a 1 mL glass bottle and seal with tinfoil;

[0080] b. After standing at room temperature for 30 minutes, add 150 μL of -SH-modified substrate probe (Probe4) with a concentration of 30 μM, mix well, and place at 4 °C for 24 hours;

[0081] c. Slowly add 50 μL of PBS buffer several times, add magnets (soaked in aqua regia the day before) and stir for 10 min, then continue to add 27 μL of PBS buffer. Take out the magnet and place it at 4°C for 48 h;

[0082] d. Transfer the labeled gold nanoparticles solution to a centrifuge tube, add sterilized water to 1 mL, centrif...

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Abstract

The invention relates to the technical field of biosensors and in particular relates to a colorimetric biosensor for detecting mercury ions and a preparation method thereof. The colorimetric biosensor is prepared from the preparation method comprising the following steps: synthesizing gold nanoparticles; modifying Probe4 on surfaces of the gold nanoparticles; mixing a marked nano gold solution and a homogeneous reaction solution. According to the colorimetric biosensor provided by the invention, high-specificity detection on the target object mercury ions is realized by utilizing specific recognition of an aptamer and a T-Hg<2+>-T composite structure; strand displacement is utilized and circulating utilization of Probe2 and Probe3 is realized and the effect of amplifying a signal is realized. The problems of a method for detecting the mercury ions at the prior art that the specificity and sensitivity are relatively low and the cost is high are solved.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a biosensor for detecting mercury ions based on nucleic acid aptamers, and also relates to a preparation method thereof. Background technique [0002] Since the existence of some heavy metal ions has adverse effects on the water environment and human health, the monitoring of heavy metal ions in aquatic ecosystems has begun to receive widespread attention. Mercury ions are toxic heavy metal pollutants widely present in water, soil and even food. Mercury ions can damage the brain, heart, stomach, intestines and kidneys. Trace amounts of mercury can accumulate in the human body through the food chain, causing chronic mercury poisoning. [0003] Currently reported detection methods for mercury ions include atomic emission spectrometry, atomic absorption spectrometry, inductively coupled plasma mass spectrometry, electrochemical methods, etc. Expensive and other issues, it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/78G01N21/33
CPCG01N21/33G01N21/78G01N33/5308
Inventor 李慧韩聪宋晓蕾刘素王玉黄加栋裴倩倩冷雪琪崔雪珺张雪王敬锋王海旺
Owner UNIV OF JINAN
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