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 o

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
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Example Embodiment

[0045] Example 1 Construction of an ultra-sensitive detection kit for mercury ions

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

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

[0048]

[0049] Among them, 2 and 2* are complementary and constitute the stem region of the stem-loop structure; 4 is the loop region of the stem-loop structure; region 1 is used to initiate the DNA strand replacement 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 auxiliary DNA is formed by forming T-Hg 2+ The -T complex binds to the 1 region of the stem-loop structure DNA, thereby starting to mediate the DNA strand replacement reaction, opening the stem-loop structure, and combining the 2* region of the auxiliary DNA with the 2 region of the stem-loop structure DNA to form a longer part Complementary double-stranded DNA. In a...

Example Embodiment

[0065] Example 2

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

[0067] (1) First use Tris-Ac buffer (20mM, containing 50mM NaCl, 10mM Mg(Ac) 2 , PH7.4) separately dissolve the stem-loop structure DNA and auxiliary DNA.

[0068] (2) Mix 500nM stem-loop structure DNA and 300nM auxiliary 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 auxiliary DNA forms T-Hg 2+ The -T complex binds to the 1 region of the stem-loop structure DNA, and starts to mediate the DNA strand replacement reaction, opens the stem-loop structure, and combines the 2* region of the auxiliary DNA with the 2 region of the stem-loop structure DNA to form a longer partial complement. Double-stranded DNA (1* and 1 complementary, 2* and 2 complementary). In addition, the 3'end of the stem-loop structure DNA forms a blunt end.

[0069] (3) Add 30U E...

Example Embodiment

[0071] Example 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, and store at room temperature.

[0074] The method of Example 2 was used to detect mercury ion solutions of different concentrations. The reaction solution (50 μL) was dropped onto the sample pad of the test strip and tested at room temperature. After 10 minutes, observe the detection area and quality control area of ​​the test strip. Color changes.

[0075] From image 3 It can be seen from the test results that when 10pM mercury ions are present, a clear red area can be observed in the detection area of ​​the test strip. The detection limit is 10pM, which is far lower than the maximum mercury ion 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 detec...

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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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