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Method for using serum SHBG as serum markers of pulmonary tuberculosis patients and application thereof

A technology for patient serum and pulmonary tuberculosis, applied in biological testing, chemiluminescence/bioluminescence, and analysis through chemical reactions of materials, etc., can solve the problems of low detection rate, long incubation time, drug resistance, etc., and achieve reliable results Effect

Active Publication Date: 2016-01-06
GUANGXI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] The purpose of the present invention is to provide a serum SHBG protein as a serum marker for tuberculosis patients and its application, to solve the problems of low detection rate and / or long culture time in the prior art, and at the same time solve the problems of current clinical anti-tuberculosis drug treatment. Drug resistance, liver function damage, gastrointestinal discomfort, allergies and other side effects, as well as poor ability of anti-tuberculosis drugs to repair lesions and sequelae

Method used

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  • Method for using serum SHBG as serum markers of pulmonary tuberculosis patients and application thereof
  • Method for using serum SHBG as serum markers of pulmonary tuberculosis patients and application thereof
  • Method for using serum SHBG as serum markers of pulmonary tuberculosis patients and application thereof

Examples

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Effect test

Embodiment 1

[0016] Increased expression of five peptides of SHBG protein in serum of tuberculosis patients detected by serum quantitative proteomics

[0017] 1. Test samples: 10 cases of multidrug-resistant tuberculosis, smear-negative tuberculosis, smear-positive tuberculosis, pneumonia patients and normal control serum. Collect 2mL whole blood on an empty stomach in the morning, let it stand at 4°C for 1-2h until the blood coagulates and precipitate the serum, centrifuge at 3000g for 10min, collect the supernatant, aliquot it on ice and store it at -80°C for later use.

[0018] 2. Detection method: (1) Removal of high-abundance proteins: According to the operation instructions of the human14 chromatographic column of the multiple affinity removal system, the high-abundance proteins in serum were removed, and the collected fractions were concentrated with a lyophilizer. (2) Desalting and detection of protein content: use 3000MWCO ultrafiltration centrifuge tube to remove high-abundance p...

Embodiment 2

[0021] Antibodies to SHBG protein containing 5 peptides including IALGGLLFPASNLR, QAEISASAPTSLR, LPLVPALDGCLR, SCDVESNPGIFLPPGTQAEFNLR, WHQVEVK were prepared, and ELISA was used to detect the expression level of SHBG protein in the serum of tuberculosis patients.

[0022] 1. Samples: Serums from 24 healthy controls, 15 pneumonia patients and 160 tuberculosis patients were collected, including 35 cases of MDR, 70 cases of smear-negative tuberculosis, and 55 cases of smear-positive tuberculosis.

[0023] 2. Detection method: ① Adding samples: set up blank wells, no samples and enzyme-labeled reagents are added to the blank control wells, and the rest of the steps are the same, standard wells, sample wells to be tested. First add 100 μl of sample diluent RD1-75 to each well of the enzyme-coated plate, and then add 50 μl of blank control, standard solution and test sample to the bottom of the well of the enzyme-labeled plate. The final dilution of the sample is 100 times. Try not ...

Embodiment 3

[0026] The primary antibody to SHBG protein containing 5 peptides including IALGGLLFPASNLR, QAEISASAPTSLR, LPLVPALDGCLR, SCDVESNPGIFLPPGTQAEFNLR, WHQVEVK was prepared, and the primary antibody was used for immunohistochemical detection, and it was found that SHBG protein was highly expressed in pulmonary tuberculosis tissue.

[0027]1. Samples: The expression level of SHBG was detected by USBiomax commercial tissue chip, which included 40 cases of pulmonary tuberculosis tissues and 10 cases of control tissues. The control tissues were normal lung tissue and lung cancer tissue.

[0028] 2. Experimental steps: ① Dewaxing: put the slide in xylene-xylene-100% alcohol-100% alcohol-95% alcohol-90% alcohol-80% alcohol-70% alcohol in sequence. Put in the first two reagents for 10 minutes, and the last six reagents for 5 minutes. ②Antigen retrieval: Rinse in clean water for a period of time after dewaxing, add 3% H 2 o 2 Soak for 10min, then pour off the H 2 o 2 , wash twice in cle...

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Abstract

The invention discloses a method for using serum SHBG (Sex hormone-binding globulin) as serum markers of pulmonary tuberculosis patients and an application thereof. The expression levels of the serum SHBG in serum of the smear negative pulmonary tuberculosis patients, the smear positive pulmonary tuberculosis patients and the multi-drug resistant pulmonary tuberculosis patients are obviously increased, detection can be carried out with the iTRAQ and MALDI-TOF / MS combined technology, mass spectrometric detection shows that the expression levels of five peptide fragments of the SHBG in the serum of the smear negative pulmonary tuberculosis patients, the smear positive pulmonary tuberculosis patients and the multi-drug resistant pulmonary tuberculosis patients are obviously higher than the expression levels in the serum of pneumonia patients and the normal, and ELISA and immunohistochemistry also verify that the SHBG is highly expressed in the serum and tissues of the pulmonary tuberculosis patients. The method and the application are suitable for auxiliary detection of pulmonary tuberculosis serum, pathogenesis researching and tuberculosis resisting drug development.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the application of a serum marker, in particular to a serum SHBG protein as a serum marker for tuberculosis patients and its application. Background technique [0002] Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis infection, with an incubation period of 4 to 8 weeks, 80% of which occur in the lungs. my country is one of the 22 countries with a high burden of tuberculosis in the world. The number of patients is second only to India, ranking second in the world. The proliferation of drugs will return TB control to the era of no chemotherapy, and about two-thirds of TB patients in the world today are at risk of developing multidrug resistance. Therefore, tuberculosis and drug-resistant tuberculosis have become serious public health and social problems in my country. Due to the current routine clinical methods of sputum smear and sputum culture to detect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N30/02G01N21/76
CPCG01N21/76G01N30/02G01N33/6884G01N2030/027
Inventor 何敏李翠萍何晓李洪涛臧宁
Owner GUANGXI MEDICAL UNIVERSITY
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