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A sodium-ion-cutting general-purpose barrier ultrafast amplification visualization sensor

A technology of sodium ion and detection method, applied in the field of biosensors, can solve the problems of cumbersome operation, long time consumption, etc.

Active Publication Date: 2020-10-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are also corresponding reports on the research results of using these methods to determine the content of sodium ions, but the operation is cumbersome, time-consuming, and requires professional personnel to operate

Method used

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  • A sodium-ion-cutting general-purpose barrier ultrafast amplification visualization sensor
  • A sodium-ion-cutting general-purpose barrier ultrafast amplification visualization sensor
  • A sodium-ion-cutting general-purpose barrier ultrafast amplification visualization sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Example 1 The establishment of a sodium ion-cutting universal partition ultrafast amplification visualization sensor

[0099] 1. Experimental materials

[0100] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, sodium chloride, 2,2-azino-bis(3-ethyl- Benzothiazole-6-sulfonic acid) diamine salt (ABTS), H2O2, 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), MES, LiCl, Tris, disodium edetate, tetramethylbis Aniline and urea were purchased from Thermo Scientific Life Technologies. All experimental water was from Milli-Q pure water system.

[0101] The sequence design is as follows (SEQ ID NO:1-7):

[0102]

[0103]

[0104] Note: The ribozyme cuts the target product and the 3' end sequence of the amplified template is complementary;

[0105] The bold base sequence added to the end of the ribozyme substrate chain is to increase the Tm value of binding to the template;...

Embodiment 2

[0126] The specificity investigation of embodiment 2 sensor

[0127] According to the biosensor constructed in Example 1, 100nM Na + , 10μM Ag + , Mg 2+ , Zn 2+ 、Cd 2+ , Hg 2 + 、Cr 3+ Added to the system for detection, the results show that the established Na + The biosensor has good specificity ( Figure 5 ).

Embodiment 3

[0128] Embodiment 3 standard addition experiment

[0129] Get high-purity water and detect with the biosensor constructed in embodiment 1, Na + 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.

[0130] Table 1 Na + Spike recovery test results

[0131]

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Abstract

The invention provides a sodium ion cutting type general partition ultrafast amplification visual sensor which comprises (1) a cutting system; (2) a sPCR (Polymerase Chain Reaction) amplication system; (3) an HCR system; (4) a detection system comprising an ABTS developing solution. According to the visual sensor disclosed by the invention, by ingeniously designing primers, templates and probes, the templates can be subjected to ultrafast amplification in the presence of sodium ions, and the amplification product enables the product to form G-quadruplex in a suitable environment by promoting the HCR reaction. Furthermore, developing is performed by utilizing mimetic peroxidase activity of the G-quadruplex, the problem that the traditional PCR product is difficult in visual detection is solved, and rapid and visual detection of the sodium ion is realized. Moreover, the sensor provided by the invention has the characteristics of high specificity and high sensitivity on the sodium ions, and the detection result is objective and accurate.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a sodium ion-cutting general-purpose barrier ultrafast amplification visualization sensor. Background technique [0002] Sodium is a metal element, the chemical symbol is Na, the atomic number is 11, and the atomic weight is 22.9898. It belongs to Group IA and is a representative of alkali metal elements. It has a soft texture and can react with water to form sodium hydroxide and release hydrogen. Chemical properties Livelier. Sodium element is widely distributed in land and ocean in the form of salt, and sodium is also one of the important components in human muscle tissue and nerve tissue. The intake of sodium in the human body is mainly through intake of food salt. Without it, the diet will be tasteless and feel weak; but if the intake is too much for a long time, it will easily affect health and induce diseases, such as making the protein content in urine Elevated, causi...

Claims

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Application Information

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