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A room temperature phosphorescence detection method and application of lysozyme

A detection method, room temperature phosphorescence technology, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of high cost, large interference, and complicated detection process, and achieve the effect of mild synthesis conditions, simple detection method, and simple preparation steps

Active Publication Date: 2021-03-30
SHANGHAI E K M BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problems of complex detection process, high cost and large interference of existing lysozyme, the present invention provides a room temperature phosphorescence detection method and application of lysozyme

Method used

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  • A room temperature phosphorescence detection method and application of lysozyme
  • A room temperature phosphorescence detection method and application of lysozyme
  • A room temperature phosphorescence detection method and application of lysozyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1, room temperature phosphorescence detection of lysozyme in urine

[0035] The first step is to prepare Mn:ZnS quantum dots:

[0036] 6-SH-β-cyclodextrin, ZnSO 4 and Mn(Ac) 2 Mix according to the molar ratio of 3:1:0.03, adjust the pH value of the system to 11 with NaOH, protect with nitrogen, and stir magnetically at room temperature for 30 minutes; then use a syringe to quickly add ZnSO 4 Equimolar Na 2 S, continue to react at room temperature for 40 min; then heat the solution to 60° C., and age in air for 3 h to obtain a crude product of Mn-doped ZnS quantum dots coated with 6-SH-β-cyclodextrin. Sediment the quantum dots with the same volume of absolute ethanol, centrifuge at high speed, pour off the supernatant, and dry in vacuum at room temperature for 24 h to obtain the desired solid powder of quantum dots.

[0037] The second step is to prepare Mn:ZnS quantum dot mother liquor:

[0038] Weigh 50mg of Mn:ZnS quantum dots, and dilute it in a 100mL vo...

Embodiment 2

[0048] Example 2, room temperature phosphorescence detection of lysozyme in urine

[0049] The first step is to prepare Mn:ZnS quantum dots:

[0050] 6-SH-β-cyclodextrin, ZnSO 4 and Mn(Ac) 2 Mix according to the molar ratio of 3:1:0.04, adjust the pH value of the system to 11 with NaOH, protect with nitrogen, and stir magnetically at room temperature for 30 minutes; then use a syringe to quickly add ZnSO 4 Equimolar Na 2 S, continue to react at room temperature for 20 min; then heat the solution to 50° C., and age in air for 2 h to obtain a crude product of 6-SH-β-CD-coated Mn:ZnS quantum dots. Sediment the quantum dots with the same volume of absolute ethanol, centrifuge at high speed, pour off the supernatant, and dry in vacuum at room temperature for 24 h to obtain the desired solid powder of quantum dots.

[0051] The second step is to prepare Mn:ZnS quantum dot mother liquor:

[0052] Weigh 50mg of Mn:ZnS quantum dots, and dilute it in a 100mL volumetric flask with s...

Embodiment 3

[0062] Example 3, room temperature phosphorescence detection of lysozyme in urine samples

[0063] The first step is to prepare Mn-doped ZnS quantum dots

[0064] 6-SH-β-cyclodextrin, ZnSO 4 and Mn(Ac) 2 Mix according to the molar ratio of 3:1:0.03, adjust the pH value of the system to 11 with NaOH, protect with nitrogen, and stir magnetically at room temperature for 30 minutes; then use a syringe to quickly add ZnSO 4 Equimolar Na 2 S, continue to react at room temperature for 20 min; then heat the solution to 50° C., and age in air for 2 h to obtain a crude product of 6-SH-β-CD-coated Mn:ZnS quantum dots. Sediment the quantum dots with the same volume of absolute ethanol, centrifuge at high speed, pour off the supernatant, and dry in vacuum at room temperature for 24 h to obtain the desired solid powder of quantum dots. The morphology, particle size and dispersion of the prepared 6-SH-β-CD-coated Mn-doped ZnS quantum dots were observed and analyzed by transmission electr...

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Abstract

The invention discloses a room-temperature phosphorescence detection method of lysozyme and an application, belongs to the technical field of detection of lysozyme, and can solve the problems of complex detection process, high cost and strong interference of lysozyme at present. A prepared Mn:ZnS room-temperature phosphorescence quantum dot modified with beta-cyclodextrin is used as a phosphorescence probe, a lysozyme nucleic acid aptamer is used as a molecular recognition unit, and analysis and detection of the lysozyme can be realized by detecting the room-temperature phosphorescence intensity of a system after the lysozyme is added. The response range of the phosphorescence detection system to the lysozyme is 5.5 nM-44.4 nM and the detection limit is 0.54 nM. The method can be used fordetecting lysozyme in serum, urine samples, honey and wine, and the complex sample pretreatment process during detection can be avoided. A deoxidant and an inducer are not required to be added for production of room-temperature phosphorescence, and interference of background fluorescence and scattered light of actual samples can be avoided.

Description

technical field [0001] The invention belongs to the technical field of detection of lysozyme, and in particular relates to a room temperature phosphorescence detection method and application of lysozyme. Background technique [0002] Lysozyme is widely found in animal body fluids, poultry protein, plants and microorganisms. Lysozyme can hydrolyze mucopolysaccharides in pathogenic bacteria, and also has many pharmacological effects such as antibacterial, anti-inflammatory, antiviral, and immune enhancement. Because of its easy digestion, easy absorption and no side effects, it has been widely used in medicine, food, bioengineering and other aspects. Clinically, lysozyme has been used to treat chronic rhinitis, acute and chronic pharyngitis, otitis media, oral ulcers and other diseases. It is also widely used in the manufacture of toothpaste and mouthwash, which can effectively prevent the occurrence of dental caries. In addition, the level of lysozyme in human body fluids c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/64
Inventor 卫艳丽左力翔兰艺凤宋秀丽李欢欢董川
Owner SHANGHAI E K M BIOTECH
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