Mercury ion detection method assisted by mimic enzyme and kit

A detection kit and detection method technology, applied in the field of biochemistry

Active Publication Date: 2021-06-08
XIANGTAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to solve the problems faced in the existing detection p

Method used

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  • Mercury ion detection method assisted by mimic enzyme and kit
  • Mercury ion detection method assisted by mimic enzyme and kit
  • Mercury ion detection method assisted by mimic enzyme and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: a kind of mercury ion detection method of the present invention, see figure 1 , when mercury ions are present, the auxiliary probe P and the hairpin probe H0 due to the formation of T-Hg 2+-T structure, thereby releasing the DNAzyme enclosed in the H0 loop of the hairpin probe. At the same time, the auxiliary probe P and the non-complementary part of the hairpin probe H0 form a split-type DNAzyme, and the two DNAzymes are joined by magnesium ions At the same time, it catalyzes the cleavage of the hairpin probe H1 of the modified ribonuclease (rA), releasing a large number of G-rich sequences, and the released G-rich sequences form G-quadruplexes in the presence of hemin and potassium ions Somatic DNase. The G-quadruplex DNase can catalyze H 2 o 2 The chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) mediated produces a blue product, which is 2 SO 4 Termination forms a stable yellow solution. The absorbance at 450 nm was measured by a UV spec...

Embodiment 2

[0047] Embodiment 2: catalytic cracking effect verification

[0048] In order to verify that the double DNAzyme design has a better catalytic cleavage effect, a control probe P0 with only one DNAzyme after opening H0 was designed. The sequence of the control probe P0 is shown in Table 1 as SEQ ID No.4. The result is as figure 2 As shown, the reaction system of the double DNAzyme has greater absorbance than the single DNAzyme, and the absorbance of the two is significantly stronger than that of the blank group, indicating that the design of the double DNAzyme has a better catalytic cracking effect.

Embodiment 3

[0049] Embodiment 3: Hairpin probe H1 and H0 concentration ratio optimization

[0050] In order to obtain the best experimental conditions, the effect of the concentration ratio of hairpin probes H1 and H0 on the absorbance was studied. The concentration of fixed auxiliary probe P and hairpin probe H0 was 20nM, and 10, 20, 40, 60, 80, 100nM hairpin probe H1 was added to the detection system respectively. According to the steps in Example 1, other conditions were kept constant. Change the concentration of hairpin probe H1 to optimize. The result is as image 3 As shown, the effect is best when the concentration of hairpin probe H1 is 60 nM. Therefore, the concentration ratio of hairpin probes H1 and H0 was selected as 3:1 as the optimal concentration ratio.

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Abstract

The invention provides a mercury ion detection method using DNAzyme to assist amplification to generate signals and a kit. The detection method comprises an auxiliary probe P, a hairpin probe H0 and a hairpin probe H1. The auxiliary probe P and the hairpin probe H0 are combined to form two independent DNAzyme in the presence of mercury ions, the hairpin probe H1 of modified ribonuclease (rA) is subjected to catalytic cracking, and a large amount of desoxyribonucleic acid sequences rich in guanine (G) are released. In hemin (hemin) and a potassium ion solution, the released sequence forms a G-quadruplex DNA enzyme to catalyze a H2O2 mediated chromogenic substrate 3, 3', 5, 5'-tetramethyl benzidine for color development, and H2SO4 is used for termination, so that a stable yellow solution is formed. The method is used for detecting mercury ions in drinking water, has the characteristics of high sensitivity, strong specificity, low cost, simplicity in operation and the like, and has a wide application prospect in actual sample detection.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a mercury ion detection method and a kit. Background technique [0002] Water-soluble mercury ion is a common inorganic form of mercury pollution, and it can cause serious harm to human health and the environment at low concentrations. For example, mercury ions ingested into the body continue to accumulate, thereby damaging DNA, inhibiting ligand-receptor interactions, and destroying the immune system, causing many serious health problems such as brain damage, kidney failure, and various cognitive and motor disorders. Limb deformation, difficulty breathing and even death may occur. Therefore, the detection of mercury ions is crucial to address environmental and health concerns. [0003] At present, atomic fluorescence spectrometry, atomic absorption spectrometry, inductively coupled plasma atomic emission spectrometry and inductively coupled plasma mass spectro...

Claims

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Application Information

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IPC IPC(8): G01N21/33G01N21/78G01N21/01
CPCG01N21/33G01N21/78G01N21/01Y02A50/30
Inventor 李文山吕育雄夏怀月孙盈盈刘文杰于建娜敬国兴刘文
Owner XIANGTAN UNIV
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