A cadmium ion-cutting general-purpose barrier ultrafast amplification visualization sensor
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A cadmium ion and sensor technology, applied in the field of biosensors, can solve the problems of poor repeatability, detection needs to be carried out in organic solvents, limitations, etc.
Active Publication Date: 2020-05-22
CHINA AGRI UNIV
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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
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Embodiment 1
[0080] Example 1 Establishment of cadmium ion-cutting universal partition ultrafast amplification visualization sensor
[0081] 1. Experimental materials
[0082] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, cadmium chloride, 2,2-azino-bis(3-ethyl- Benzothiazole-6-sulfonic acid) diamine salt (ABTS), H2O2, potassium chloride, sodium chloride, magnesium chloride, potassium hydrogen phosphate, disodium edetate, urea, 4-hydroxyethylpiperazine Sulfonic acid (HEPES), sodium hydroxide, disodium hydrogen phosphate were purchased from Thermo Scientific Life Technologies. All experimental water was from Milli-Q pure water system.
[0083] The sequence design is as follows (SEQ ID NO:1-5):
[0084]
[0085] Note: The ribozyme cuts the target product and the 3' end sequence of the amplified template is complementary;
[0086] The bold base sequence added to the end of the ribozyme su...
[0106] According to the biosensor constructed in Example 1, 100nM Cd 2+ , 10 μM Ag + , Mg 2+ , Zn 2+ 、Cd 2+ , Hg 2+ Added to the system for detection, the results show that the established Cd 2+ The biosensor has good specificity ( Figure 4 ).
Embodiment 3
[0107] Embodiment 3 standard addition experiment
[0108] Get high-purity water and detect with the biosensor constructed in embodiment 1, Cd 2+ 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.
[0109] Table 1Cd 2+ Spike recovery test results
[0110]
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Abstract
The invention provides a cadmium ion cutting type universal type isolating superfast amplification visualization sensor. The cadmium ion cutting type universal type isolating superfast amplification visualization sensor comprises 1, a cutting system, 2, a sPCR amplification system, and 3, a detection system containing ABTS color developing solution. According to a preparation method, skilful design of primers, templates, and probes is adopted, so that in the presence of cadmium ions, template superfast amplification is realized, a G-quadruplex is formed in an appropriate environment from amplification products; and the G-quadruplex peroxidase activity is adopted in color development, so that a difficult problem in the prior art that visualization detection of conventional PCR products is difficult to realize is solved, and rapid visualization detection on cadmium ions is realized. And in addition, the cadmium ion cutting type universal type isolating superfast amplification visualization sensor is high in specificity and sensitivity on cadmium ions, so that detection results are more objective and accurate.
Description
technical field [0001] The invention relates to the technical field of biosensors, in particular to a cadmium ion cutting type universal partition ultrafast amplification visualization sensor. Background technique [0002] Cadmium element symbol Cd, blue-white metal, belongs to Group IIB in the periodic table of elements, the atomic number of cadmium is 48, the atomic weight is 112.41, and the oxidation state is +1, +2. It is toxic, but due to its good flexibility and oxidation resistance , are widely used in industrial production. Cadmium can enter the organism through the respiratory and digestive tracts, and the cadmium entering the body can cause damage to a series of important organs such as the liver, kidney, bone, brain, and lung, and can also cause damage to the nervous, reproductive, and immune systems. China is the largest cadmium reserve and development country in the world, and it is also the largest cadmium consumer. In recent years, with the further developme...
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