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Kit for measuring content of lysozyme in human body fluid sample on basis of latex-enhanced immunoturbidimetric assay

A technology of human lysozyme and latex immunization, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of time-consuming and insufficient detection speed, and achieve the effects of short time-consuming, improved detection sensitivity, and improved detection efficiency

Active Publication Date: 2018-12-14
SHENYANG BAICHUANGTE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method also takes a lot of time. In practical applications, detection methods that are not fast enough will be eliminated soon.

Method used

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  • Kit for measuring content of lysozyme in human body fluid sample on basis of latex-enhanced immunoturbidimetric assay
  • Kit for measuring content of lysozyme in human body fluid sample on basis of latex-enhanced immunoturbidimetric assay
  • Kit for measuring content of lysozyme in human body fluid sample on basis of latex-enhanced immunoturbidimetric assay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Preparation of human lysozyme antibody in the kit of Example 1

[0052] 1) Prepare the human lysozyme nucleic acid sequence into cDNA, and transfer it into the yeast expression system by genetic engineering technology;

[0053] 2) Ferment the yeast at 30°C, pH 4.5, wort medium, and obtain the crude product of the fermentation liquid containing human lysozyme, mix the crude product of human lysozyme with 724 resin, stir and adsorb for 6h, and put it under the condition of 2°C Stand still for 24 hours, pour off the supernatant;

[0054] 3) Centrifuge the resin at 8000rpm for 2h, pour off the supernatant, wash the resin repeatedly with deionized water, repeat this step twice, pack the resin into a column, wash the resin with 150mM pH 6.5 phosphate buffer, and wash the resin with 10 % ammonium sulfate solution to elute the resin, and collect the eluate;

[0055] 4) Under the condition of stirring at 200rpm, slowly add solid ammonium sulfate to the eluent to make the conce...

Embodiment 2

[0058] Preparation of polystyrene microsphere preservation solution in the kit of embodiment 2

[0059] 1) Add 800mL of 50mM pH 8.0 HEPES buffer solution, 50g BSA, stir to dissolve, 9gNaCl, 1g skimmed milk powder, 1mL TWEEN-20, 1g maltose, 1mL PEG8000, add each component while stirring, and then use 50mM pH 8.0HEPES buffer volume to 1000mL;

[0060] 2) Filter the solution through a 0.22 μm microporous membrane for later use.

Embodiment 3

[0073] Preparation of calibrator in embodiment 3 kit

[0074] 1) Add 10% (W / V) BSA, 0.1% (V / V) TWEEN-20 to 20mM pH 6.0 PBS buffer solution, stir well and then filter with 0.22μm microporous membrane to prepare the calibration product dilution ;

[0075] 2) The calibrator can be obtained by dissolving human lysozyme with the diluent of the calibrator.

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Abstract

The invention belongs to the technical field of biomedical service and particularly relates to a kit for measuring content of lysozyme in a human body fluid sample on the basis of latex-enhanced immunoturbidimetric assay. The kit contains a reagent 1 and a reagent 2, wherein the reagent 1 is a reaction buffer and is specifically a 100mM PBS buffer solution containing 0.1% (V / V) TWEEN-20 and havingpH of 7.2-7.4; the reagent 2 is a solid-phase carrier linked with a human lysozyme antibody and is a specifically polystyrene microsphere. The kit can assist in clinical diagnosis, detection sensitivity for lysozyme content is improved remarkably, and a clinical method for measuring the lysozyme content is simplified.

Description

field of invention [0001] The invention belongs to the technical field of biomedical detection, and in particular relates to a kit for detecting the content of lysozyme in human body fluid samples based on latex immunoturbidimetry. Background of the invention [0002] Peptidoglycan is the main component of the bacterial cell wall, mainly connected by NAM and NAG through β-1,4 glycosidic bonds, and the peptide "tail" of NAG is connected to the third carbon atom of NAM through D-lactyl carboxyl group , the peptide tails are connected by peptide "bridges", that is, peptide bonds or a few amino acids. NAM, NAG, peptide "tails" and peptide "bridges" together form a multilayer network structure of bacterial cell wall peptidoglycan, as The skeleton of the cell wall, any breakage of chemical bonds in the above structure can cause damage to the bacterial cell wall. [0003] Lysozyme (lysozyme) is an alkaline enzyme with a molecular weight of about 15,000 and composed of about 129 am...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/573
CPCG01N33/558G01N33/573
Inventor 李伟东
Owner SHENYANG BAICHUANGTE BIOTECH
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