Unlock instant, AI-driven research and patent intelligence for your innovation.

Tuberculosis serum marker screening method and application

A technology for serum markers and screening methods, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problems of non-specificity, protein omission, non-specific binding, etc., and achieve high specificity, high protein biological activity, and accuracy. high sex effect

Pending Publication Date: 2022-03-25
NINGXIA MEDICAL UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research on serum markers of tuberculosis is a separate, non-specific research model; there are the following limitations: (1) about 50% of Mycobacterium tuberculosis (M.tuberculosis, MTB) in exogenous E. coli (E.coli) host cells ) proteins cannot be expressed, resulting in the omission of such proteins in the screening of serum markers
(2) The expression, separation and purification process of a large number of MTB recombinant proteins is time-consuming and laborious, and most of the recombinant expressed proteins are not ideal serum markers
(3) The activity of exogenously expressed MTB protein is easily affected by the nature of the expression host itself; (4) The MTB protein is recognized by the patient's total serum, because the total protein composition of the serum is very complex, and there is a great possibility of non-specific binding, so It is easy to cause false positive test results
[0003] In order to solve the limitations of the screening of tuberculosis serum markers in the prior art, an effective screening method is needed

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Tuberculosis serum marker screening method and application
  • Tuberculosis serum marker screening method and application
  • Tuberculosis serum marker screening method and application

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0045] Preparation Example 1 Constructing a Biomimetic Affinity Ligand Library

[0046] The biomimetic affinity ligand synthesis process is similar to the content of our previously published patent: "Antibody Affinity Purification Material and Its Application, 2014.11.19, China, CN104148018A".

[0047] The biomimetic affinity small molecule structures constructed this time for the specific adsorption of human serum antibodies include but are not limited to the following 19 types: L-theanine; aminopyrine; 3-aminophenol; 4-amino-1 naphthol hydrochloride ;4-2aminoethylbenzenesulfonamide;1-aminoanthraquinone;hexylamine;2,6-diaminoanthraquinone;2,6-diaminopyridine;2-phenylacetamide;adenine;hydroxysuccinyl imine; 2,4,6-triaminopyrimidine; diphenylamine; dodecylamine; diethylamine hydrochloride; dibenzylamine; dodecylamine; N-acetyl-L-methionine.

[0048] Take the bionic affinity medium A5-87 (5:4-2aminoethylbenzenesulfonamide, 87:ethylenediamine) to separate and purify the mixed se...

preparation example 2

[0050] Preparation example 2 sample pretreatment:

[0051] Experimental group: From 20 tuberculosis patient serum samples, take 100 μL per person, and obtain a total of 2 mL serum samples, centrifuge and mix at 2000 rpm, discard the precipitate, and dilute the serum 8 times with 0.005-0.5 mol of PBS buffer as it is;

[0052] Control group: take 20 healthy human serum samples, and the processing method is the same as that of the above-mentioned experimental group.

[0053] Equilibration of the bionic affinity chromatography column: wash the bionic affinity separation matrix gravity column with 10mL 0.1M acetic acid, 10mL double distilled water, 10mL 0.1M NaOH, and 10mL double distilled water in sequence, and finally wash the bionic affinity separation matrix gravity column with 10mL PBS (10mM, pH 7.4) Equilibrate all columns separately.

[0054] Biomimetic affinity chromatography of serum samples after pretreatment: take 5 mL diluted serum sample solutions of the experimental...

Embodiment 1

[0059] Such as figure 2 The process flow of the present invention is shown. The prepared Mycobacterium tuberculosis culture filtrate or cytoplasmic protein of Mycobacterium tuberculosis cells were subjected to chromatography with the bionic affinity chromatography column of the experimental group and the bionic affinity chromatography column of the control group respectively. Among them, the bionic affinity chromatography column in the experimental group and the bionic affinity chromatography column in the control group were chromatographically made of A5-87 material.

[0060] Liquid Chromatography-Mass Spectrometry Analysis of Eluted Components:

[0061] Collect all the eluted control groups and experimental histone components adsorbed on the bionic affinity chromatographic column A5-87 by the bionic-immunoaffinity method; and wash all the eluted components with 25mM NH 4 HCO 3 Replace buffer system. Then all samples were hydrolyzed with Trypsin, and the hydrolyzed polyp...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Concentrationaaaaaaaaaa
Login to View More

Abstract

The invention discloses a tuberculosis serum marker screening method which comprises the following steps: respectively carrying out bionic affinity or affinity chromatography on serum (experimental group) of a tuberculosis patient and serum (control group) of a healthy person, preparing a bionic affinity or affinity chromatography antibody column of the experimental group, and preparing a bionic affinity or affinity chromatography antibody column of the control group; respectively carrying out biomimetic affinity or affinity chromatography antibody column chromatography on the mycobacterium tuberculosis culture filtrate or mycobacterium tuberculosis cytoplasm protein and the experimental group and the control group; analyzing mycobacterium tuberculosis protein combined with serum antibodies of the experimental group and the control group to preliminarily obtain a serum marker for tuberculosis detection; and further analyzing the primarily selected tuberculosis detection serum marker by a method. The method combines bionic affinity chromatography, antigen-antibody specific recognition, mass spectrometry and bioinformatics analysis methods to realize high-throughput screening and evaluation of serum markers of tuberculosis patients. The limitation of a tuberculosis serum marker screening method in the prior art is solved.

Description

technical field [0001] The invention relates to the field of medical diagnosis, in particular to a screening method for tuberculosis serum markers and the application of the method in the field of tuberculosis serum diagnosis. Background technique [0002] Tuberculosis (TB) is one of the infectious diseases with the highest single morbidity and mortality rate caused by Mycobacterium tuberculosis (MTB) infection body, and pulmonary tuberculosis patients account for 90% of the total number of patients. The current research on serum markers of tuberculosis is a separate, non-specific research model; there are the following limitations: (1) about 50% of Mycobacterium tuberculosis (M.tuberculosis, MTB) in exogenous E. coli (E.coli) host cells ) proteins cannot be expressed, resulting in the omission of such proteins in the screening of serum markers. (2) The expression, separation and purification process of a large number of MTB recombinant proteins is time-consuming and labori...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 马国荣徐锐强杨延辉马锐李荣秀王佩杨玉荣罗鹏征朱娜
Owner NINGXIA MEDICAL UNIV