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Ultra-sensitive detection method of mercury ions and detection kit

A technology of sensitive detection and detection method, applied in the field of analytical chemistry, can solve the problems of low detection sensitivity, difficult to meet the needs of environmental sample detection, etc., and achieve the effects of high sensitivity, intuitive detection results, and convenient detection process.

Active Publication Date: 2014-05-07
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the potential disadvantage is that the detection sensitivity is low, and it is difficult to meet the needs of environmental sample detection, so it is necessary to improve the detection sensitivity through signal amplification

Method used

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  • Ultra-sensitive detection method of mercury ions and detection kit
  • Ultra-sensitive detection method of mercury ions and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The construction of a kind of ultrasensitive detection kit of mercury ion of embodiment 1

[0046] 1. Design of stem-loop DNA and auxiliary DNA

[0047] The sequence of the stem-loop DNA is shown in SEQ ID NO: 1, and its structure is as follows:

[0048]

[0049] Among them, 2 and 2* are complementary, constituting the stem-like region of the stem-loop structure; 4 is the loop region of the stem-loop structure; and 1 region is used to initiate the DNA strand displacement reaction.

[0050] The sequence of the auxiliary DNA is shown in SEQ ID NO: 2, and the structure is as follows:

[0051]

[0052] The 1* region of the helper DNA passes through the formation of T-Hg 2+ The -T complex binds to the 1 region of the stem-loop structure DNA, thereby initiating the DNA strand displacement reaction, opening the stem-loop structure, allowing the 2* region of the auxiliary DNA to combine with the 2 region of the stem-loop structure DNA to form a longer part Complementar...

Embodiment 2

[0066] Utilize the test kit constructed in Example 1 to detect samples, the steps are as follows:

[0067] (1) Use Tris-Ac buffer (20mM, containing 50mM NaCl, 10mM Mg(Ac) 2 , pH7.4) to dissolve the stem-loop structure DNA and the auxiliary DNA respectively.

[0068] (2) Mix 500nM stem-loop DNA and 300nM helper DNA, then add the sample to be tested, mix well, and place at room temperature for 30 minutes. At this step, if there is Hg in the sample to be tested 2+ , the 1* region of the helper DNA passes through the formation of T-Hg 2+ The -T complex binds to the 1 region of the stem-loop structure DNA, and starts to mediate the DNA strand displacement reaction, opening the stem-loop structure, allowing the 2* region of the auxiliary DNA to combine with the 2 region of the stem-loop structure DNA to form a longer partial complementarity Double-stranded DNA (1* and 1 are complementary, 2* and 2 are complementary). Also, the 3' end of the stem-loop DNA forms a blunt end.

[0...

Embodiment 3

[0072] Detection of different concentrations of mercury ions:

[0073] Prepare mercury ion standard solutions with concentrations of 10pM, 100pM, 1nM, 10nM, 100nM and 500nM respectively, and store at room temperature.

[0074] Adopt the method of embodiment 2 to detect the mercury ion solutions of different concentrations, drip the reaction solution (50 μ L) on the sample pad of the test paper, detect at room temperature, observe the detection area of ​​the test paper and the quality control area after 10 minutes Color changes.

[0075] From image 3 It can be seen from the test results that when 10pM mercury ions exist, a clear red area can be observed in the detection area of ​​the test strip, and its detection limit is 10pM, which is far lower than the maximum limit of mercury ions in drinking water stipulated by the US Environmental Protection Agency. The allowable amount is 10nM. And, as the concentration of mercury ions increases (from 10pM to 100nM), the color of the...

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PUM

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Abstract

The invention discloses an ultra-sensitive detection method of mercury ions and a detection kit. According to the ultra-sensitive detection method, an exonuclease III mediated signal amplification technology and a test strip detection technology are combined so that a quick determination method of trace mercury ions is constructed. The ultra-sensitive detection method is very high in sensitivity; the detection limit to mercury ions is 10 pM, a detection result is visual and macroscopic, the detection process is convenient and rapid, and the ultra-sensitive detection method can be used in on-site field testing of mercury ions; in addition, the detection method disclosed by the invention is better in specificity, and other common metal ions have no influence to the detection; and the constructed test strip is time-saving and labor-saving in use, high in detection speed, simple and convenient to operate, visual and macroscopic in detection result, and free from using any instrument.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a rapid detection method for trace mercury ions by signal amplification mediated by exonuclease III. Background technique [0002] Mercury ion (Hg 2+ ) is a ubiquitous heavy metal pollution source, which is seriously harmful to the human body and is an important indicator for environmental monitoring. Exposure to mercury ions can cause a range of adverse health effects in varying degrees, including kidney failure, brain damage, nervous system and immune system damage. The United States Environmental Protection Agency stipulates that the maximum allowable amount of mercury ions in drinking water shall not exceed 10nM. At present, the routine detection methods of mercury ions mainly include atomic absorption method, atomic fluorescence spectrometry, inductively coupled plasma emission spectrometry and so on. However, these methods are cumbersome to opera...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/682C12Q2525/185C12Q2521/325C12Q2565/625
Inventor 陈俊华周顺桂温俊林
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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