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SHISA4 serving as biomarker for early diagnosis of chronic obstructive pulmonary disease

A chronic obstructive pulmonary and material technology, applied in disease diagnosis, biological testing, biomaterial analysis, etc., can solve problems such as clinical misdiagnosis, and achieve the effect of improving survival rate

Inactive Publication Date: 2018-05-15
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, more and more evidence has found that using FEV1 / FVC<0.70 as the diagnostic standard of COPD can lead to a large number of clinical misdiagnosis, and the current diagnostic standard is facing unprecedented challenges

Method used

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  • SHISA4 serving as biomarker for early diagnosis of chronic obstructive pulmonary disease
  • SHISA4 serving as biomarker for early diagnosis of chronic obstructive pulmonary disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Screening of differentially expressed genes in patients with chronic obstructive pulmonary disease and normal people

[0042] 1. Clinical research objects:

[0043] Five patients with chronic obstructive pulmonary disease were selected, including 2 males and 3 females, ranging in age from 50 to 79 years old. The diagnostic criteria were in compliance with the "Chronic Obstructive Pulmonary Disease Diagnosis and Treatment Norms" revised in 2007.

[0044] Diagnostic criteria: Any patient suffering from dyspnea, chronic cough or sputum, and a history of exposure to risk factors, lung function tests show that after inhalation of bronchodilators, there is airflow limitation, which can be diagnosed as COPD.

[0045] Exclusion criteria: ①Patients with other lung diseases, such as bronchial asthma, pulmonary fibrosis, lung cancer, etc.; ②Patients with other infections; ③With severe cardiovascular and cerebrovascular diseases, diabetes, blood system diseases, malignant tumors, ...

Embodiment 2

[0085] Example 2 QPCR verification of the relationship between candidate genes and chronic obstructive pulmonary disease

[0086] Based on the results of previous high-throughput sequencing and according to the size of P value, we selected SHISA4 gene (its expression is down-regulated in patients with chronic obstructive pulmonary disease) for verification.

[0087] 1. Research objects:

[0088] According to the method of Example 1, 45 patients with chronic obstructive pulmonary disease and 35 normal persons were selected.

[0089] 2. Extraction of total RNA from blood

[0090] Use Biotech blood RNA extraction kit to extract total RNA from blood:

[0091] (1) Take 250μl (or 0.25g) of whole blood into the RNase-Free filter column, centrifuge at 13000rpm for 2 minutes, collect the lower liquid, and add 0.75ml lysis buffer RLS.

[0092] (2) The homogenized sample was shaken vigorously and mixed, and incubated at 15-30°C for 5 minutes to completely decompose the ribosome.

[0093] (3) Optional...

Embodiment 3

[0109] Example 3 Western blotting experiments to verify the expression products of differentially expressed genes in patients with chronic obstructive pulmonary disease and normal people

[0110] 1. Clinical object: Same as Example 2.

[0111] 2. Separation of monocytes

[0112] For patients with chronic obstructive pulmonary disease and normal people, take 10ml of venous blood, inject it into a sterile vial containing heparin, and shake it gently after capping. Use a sterile pipette to add an equal volume of HBSS (NaCl 8.0g, Na 2 HPO 4 0.132g, KH 2 PO 4 0.06g, KCl 0.4g, phenol red 1ml, NaHCO 3 0.35g, D-glucose 1.0g, dissolved in 1000ml double distilled water) to reduce the aggregation of red blood cells. Aspirate 8ml of lymphocyte layering solution and place it in a 50ml centrifuge tube. Add diluted blood slowly along the tube wall to keep the interface clear. Do not mix the two. Centrifuge at 2000r / min for 30min at 20℃. Carefully draw the layered solution and connect it with plas...

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Abstract

The invention discloses an SHISA4 gene serving as a diagnosis tool for a chronic obstructive pulmonary disease. High-throughput sequencing and QPCR (Quantitative Polymerase Chain Reaction) researchesfind out that the content of the SHISA4 gene in blood of a patient with the chronic obstructive pulmonary disease is remarkably reduced when being compared with the content in a normal person; a western blot experiment proves that the content of SHISA4 protein in the blood of the patient with the chronic obstructive pulmonary disease is remarkably reduced when being compared with the content in the normal person. Therefore, the SHISA4 gene and the SHISA4 protein can be used as biomarkers for diagnosing the chronic obstructive pulmonary disease. A method for characterizing the chronic obstructive pulmonary disease by utilizing the content of the gene or the protein in the blood has the following advantages that tissue sampling detection is avoided and pains of subjects are alleviated; meanwhile, the characterizing method can be used for screening healthy people and the chronic obstructive pulmonary disease is effectively prevented.

Description

Technical field [0001] The present invention relates to the field of chronic obstructive pulmonary disease diagnosis. More specifically, the present invention relates to SHISA4 gene as a diagnostic tool for chronic obstructive pulmonary disease. Background technique [0002] Chronic obstructive pulmonary disease (COPD) is a common and frequently-occurring disease in the respiratory department. It is also an internationally recognized chronic pulmonary disease that seriously threatens people's health. Its morbidity and mortality are high, and it seriously increases the economic burden of people and society. According to statistics from the World Health Organization, COPD was the fourth leading cause of death in the world in 2000 (Aibek E Mirrakhimov. Chronicobstructive pulmonary disease and glucose metabolism: a bitter sweetsymphony. Cardiovasc Diabetol, 2012, 11: 132). It is expected that by 2020, COPD will be It has risen to the third leading cause of death and the fifth leading...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/68G01N2800/122
Inventor 向常娟马翠
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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