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Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for chromium and cadmium

A technology of amplification system and cleavage site, applied in the field of biosensors, can solve the problems of poor repeatability, detection in organic solvents, limitations, etc.

Active Publication Date: 2018-12-07
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods rely on large-scale instruments and special personnel to operate, and are limited in field and field detection
Chemical sensing methods represented by small organic molecules such as ethers, polyamines, and aromatic hydrocarbons have also been developed to a certain extent, but at present there are shortcomings such as low sensitivity, poor repeatability, and the detection needs to be carried out in organic solvents. Reliability and usability are not high

Method used

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  • Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for chromium and cadmium
  • Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for chromium and cadmium
  • Universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for chromium and cadmium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0122] Example 1 Chromium- and cadmium-cut functional nucleic acid visual detection method for universal partition ultrafast amplification

[0123] 1. Experimental materials

[0124] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, chromium chloride, cadmium chloride, 2,2-azino-bis(3 -Ethyl-benzothiazole-6-sulfonic acid) diamine salt (ABTS), H 2 o 2 , morpholineethanesulfonic acid monohydrate (MES), potassium chloride, sodium chloride, chromium chloride, potassium hydrogen phosphate, disodium edetate, urea, 4-hydroxyethylpiperazineethanesulfonic acid (HEPES ), sodium hydroxide, and disodium hydrogen phosphate were all purchased from Thermo Scientific Life Technologies. All experimental water was from Milli-Q pure water system.

[0125] The sequence design is as follows (SEQ ID NO:1-8):

[0126]

[0127]

[0128] Note: The ribozyme cuts the target product and the 3' end se...

Embodiment 2

[0156] The specificity investigation of embodiment 2 detection method

[0157] According to the biosensor constructed in Example 1, 1nM Cr 3+ , 100nM Cd 2+ , 10 µM Cu 2+ , Zn 2+ , Mg 2+ , Hg 2+ 、Na + 、Ag + , added to the system for detection, the results show that the established Cr 3+ The biosensor has good specificity ( Figure 7 ).

Embodiment 3

[0158] Embodiment 3 standard addition experiment

[0159] Get high-purity water and detect with the biosensor constructed in embodiment 1, Cd 2+ 、Cr 3+ No detection was carried out, and the standard addition experiment was carried out on it, and the results obtained by continuous measurement are shown in Table 1.

[0160] Table 1 The results of the standard recovery experiment

[0161]

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Abstract

The invention provides a universal-partition ultrafast-amplification cleavage type functional-nucleic acid visual detection method for chromium and cadmium. According to the invention, primers, a template and probes are ingeniously designed, so ultrafast amplification of the template can be realized in the presence of chromium and cadmium ions, and amplification products are allowed to form G-quadruplexes in a proper environment; and color development is performed by utilizing the peroxidase-like activity of the G-quadruplexes. Thus, the problem of difficulty in realizing visual detection of traditional PCR products is overcome, and rapid and visual detection of chromium and cadmium ions is realized. Moreover, the method provided in the invention has the characteristics of high specificityand high sensitivity to chromium and cadmium ions, and is more objective and accurate in detection results.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a visual detection method for chromium and cadmium cutting functional nucleic acid with universal partition ultrafast amplification. Background technique [0002] Chromium element symbol Cr, silver-white metal, belongs to group VIB in the periodic table of elements, the atomic number of chromium is 24, the atomic weight is 51.996, body-centered cubic crystal, and the common valences are +3, +6 and +2. The average content of chromium in the earth's crust is 0.010% to 0.011%, and the distribution is extremely uneven. my country's chromium resources are not abundant, and trivalent chromium is one of the indispensable trace elements for the human body. Adults need about 500-700 micrograms of chromium per day. When chromium is deficient, it is easy to make the human body suffer from diabetes and coronary atherosclerosis, which affects the fat metabolism in the body. However, too ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2521/337C12Q2525/301C12Q2565/607
Inventor 罗云波许文涛黄昆仑杜再慧田晶晶
Owner CHINA AGRI UNIV
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