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Nano biosensor method and kit for rapid detection of Hg ions

A technology of biosensors and mercury ions, which is applied in the measurement of color/spectral characteristics, analysis by making materials undergo chemical reactions, and material analysis by observing the impact on chemical indicators, etc., can solve the tedious and time-consuming problems, and the sensitivity needs to be improved. , expensive and other issues, to achieve the effect of simple operation, short oligonucleotide probe sequence and high sensitivity

Inactive Publication Date: 2012-07-25
WENZHOU MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This detection method can be carried out at room temperature, but requires three oligonucleotide probes, two of which still need to be modified with sulfhydryl groups, which is tedious, time-consuming and expensive
Lihua Wang et al. (Gold Bulletin 2008, 41, (1), 37-41) reported that a poly-T oligonucleotide probe was used to detect 2+ Specifically stabilize the T-T mismatch to form a stem-loop structure, so that the gold nanoparticles bound to the poly-T oligonucleotide probe lose protection and aggregate under high-salt conditions, and produce a red-blue color change, realizing the anti-Hg 2+ Fast and convenient analysis, but the sensitivity needs to be improved

Method used

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  • Nano biosensor method and kit for rapid detection of Hg ions
  • Nano biosensor method and kit for rapid detection of Hg ions
  • Nano biosensor method and kit for rapid detection of Hg ions

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1MSO optimal design

[0031] The MSO sequences selected in this study are shown in Table 1, where MSO 1 It is a mercury ion-specific oligonucleotide probe (stem-loop structure) reported by Lihua Wang et al. In order to improve the sensitivity of mercury ion detection and reduce the cost of DNA synthesis, MSO 1 Carry out an optimized design, remove the non-specific loop part (4bp), intercept the stem part (9bp) at both ends, and mark them as MSO 2 and MSO 3 .

[0032] Table 1 Mercury ion-specific oligonucleotides

[0033]

[0034] Take 3.5 μL MSO (10 μmol / L MSO 1 , 20μmol / L MSO 2 , 20μmol / L MSO 3 or MSO 2 and MSO 3 Each 10μmol / L mixture), add 3.5μL Hg 2+ (100μmol / L) was made up to a total volume of 20μL with ultrapure water, reacted at room temperature for 5 minutes, then added 100μL nano-gold, reacted at room temperature for 5 minutes, and finally added 13μL NaNO 3 . Observe the color change, and record the A650 and A522 of each well with a Var...

Embodiment 2

[0035] Example 2NaNO 3 Concentration optimization

[0036] Take 13 μL NaNO with different concentrations (0.25, 0.5, 1, 2mol / L) 3 , using the above-mentioned optimally designed MSO for colorimetric detection, using a Varioskan Flash multi-functional microplate reader to record the A650 and A522 of each well, and at the same time, each group uses ultrapure water as a blank control to calculate the different NaNO 3 Detection of Hg under concentration 2+ The S / N. figure 2 In we can see NaNO 3 Concentration increased from 0.25 to 0.5mol / L, nano biosensor detects Hg 2+ The S / N also increased, reaching 6.83±0.08. but continue to increase NaNO 3 , S / N decreased on the contrary, it may be that too high salt concentration will make the blank control also change in color, so the sodium nitrate of 0.5mol / L is selected as the optimal concentration.

Embodiment 3

[0037] Embodiment 3 colorimetric detection Hg 2+

[0038] We combined the mercury ion nanobiosensor with Hg 2+ (1mmol / L) and ultrapure water reaction, draw A 650 / A 522 curve over time. Such as image 3 As shown, adding NaNO 3 After just 5 seconds, the mercury ion nanobiosensor can successfully distinguish Hg 2+ and ultrapure aqueous solutions. And as time goes on, Hg 2+ (1mmol / L) group A 650 / A 522 also increases, reaching equilibrium at 15 minutes (A 650 / A 522 =1.13); while the ultrapure water group is still only about 0.16, it can be considered that there is almost no change.

[0039] In order to further analyze the detection sensitivity of the nano-biosensor, a series of different concentrations of Hg 2+ to test. The color of nano-gold begins to change at 20 μmol / L with the naked eye, and the greater the concentration, the more obvious the change; through colorimetric analysis, the lowest detectable value is 10 μmol / L (such as Figure 4 , 3 times the standa...

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Abstract

The invention relates to a nano biosensor capable of carrying out rapid colorimetric detection on Hg<2+> in water. The nano biosensor is used for carrying out optimized design on a Hg ion specificity oligonucleotide probe on the basis of the Hg ion specificity oligonucleotide probe and nanogold and adsorbing the Hg ion specificity oligonucleotide probe to the surfaces of nanogold particles to realize assembly of the biosensor; the quantitative analysis on Hg<2+> is realized through utilizing stable T-T mispairing of Hg ion specificity and color change of the nanogold particles under the high-salt condition. The nano biosensor is free of marking the probe or modifying the nanogold, can realize visible portable analysis of the Hg ions without any special condition, has the advantages of simpleness and convenience for operation, short detection time, higher sensitivity, strong specificity and the like and is easy to popularize and use.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and relates to a method for constructing a nano-biological sensor for rapid colorimetric detection of mercury ions in water by using mercury ion-specific oligonucleotide probes and nano gold and a kit thereof. Background technique [0002] The detection of mercury ions has long been of great concern to researchers. Since the industrial revolution, mercury has been used in production and life, the mercury content in the environment is increasing day by day, and the scope of mercury pollution is also expanding year by year. According to statistics, the content of mercury in global water bodies has increased by about three times compared with a few years ago, and the content of mercury near industrial areas is even higher. Due to the high mobility and persistence of mercury pollution, it can cause great harm to the environment and human health through methylation, bioaccumulation, and ...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/31
Inventor 吕建新吴文鹤
Owner WENZHOU MEDICAL UNIV
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