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Application of DEFA1B gene to preparation of tool for diagnosing chronic obstructive pulmonary disease

A chronic obstructive pulmonary disease and gene technology, applied in the field of biomedicine, can solve the problem of the overall prevalence of chronic obstructive pulmonary disease and other problems, and achieve the effect of being conducive to diagnosis

Active Publication Date: 2018-09-04
CHINA JAPAN FRIENDSHIP HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are vertical differences in the survey data, it can still be seen that the overall prevalence of COPD is grim

Method used

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  • Application of DEFA1B gene to preparation of tool for diagnosing chronic obstructive pulmonary disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Screening of differentially expressed genes between patients with chronic obstructive pulmonary disease and normal people

[0037] 1. Clinical research object:

[0038] Six patients with chronic obstructive pulmonary disease were selected, including 3 males and 3 females, with an age range of 52-76 years old. The diagnostic criteria were all in line with the "Code for Diagnosis and Treatment of Chronic Obstructive Pulmonary Disease" revised in 2007 in my country.

[0039] Diagnostic criteria: COPD can be diagnosed in any patient with dyspnea, chronic cough, or phlegm production, with a history of exposure to risk factors, and pulmonary function tests showing airflow limitation following inhaled bronchodilators.

[0040] Exclusion criteria: ①Combined with other lung diseases, such as bronchial asthma, interstitial pulmonary fibrosis, lung cancer, etc.; ②Patients with infection in other parts; organ failure, hepatitis; ④ suffering from immune system diseases or...

Embodiment 2

[0080] Example 2 QPCR verification of the relationship between candidate genes and chronic obstructive pulmonary disease

[0081] Based on the previous high-throughput sequencing results and according to the size of the P value, we selected the DEFA1B gene (its expression is upregulated in patients with chronic obstructive pulmonary disease) for verification.

[0082] 1. Research object:

[0083] According to the method of Example 1, 35 patients with chronic obstructive pulmonary disease and 30 normal people were selected.

[0084] 2. Total RNA extraction from blood

[0085] Use Biotech Blood RNA Extraction Kit to extract total RNA from blood:

[0086] (1) Take 250 μl (or 0.25 g) of whole blood into an RNase-Free filter column, centrifuge at 13,000 rpm for 2 minutes, collect the lower liquid, and add 0.75 ml of lysis solution RLS.

[0087] (2) Shake the homogenate sample vigorously, and incubate at 15-30°C for 5 minutes to completely decompose the ribosomes.

[0088] (3) O...

Embodiment 3

[0104] The preparation of embodiment 3 chronic obstructive pulmonary disease diagnostic kits

[0105] According to the correlation between DEFA1B gene and chronic obstructive pulmonary disease, whether chronic obstructive pulmonary disease occurs can be diagnosed by detecting the expression of DEFA1B gene. Accordingly, the present invention provides a method based on detecting DEFA1B gene expression to diagnose chronic obstructive pulmonary disease A disease kit, the components of the diagnostic kit are as follows: a SYBR Green polymerase chain reaction system; a pair of primers for amplifying DEFA1B gene and GAPDH gene. The forward sequence of DEFA1B gene is 5'-AAAGCATCCAGGCTCAAG-3', the reverse sequence is 5'-TTCCATAGCGACGTTCTC-3'; the forward primer sequence of GAPDH amplification is 5'-AACTCTGGTAAAGTGGATATTG-3', and the reverse primer sequence is 5' '-GGTGGAATCATATTGGAACA-3'. The SYBR Green polymerase chain reaction system includes PCR buffer, dNTPs, and SYBR Green fluore...

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Abstract

The invention discloses genes related to diagnosis of the chronic obstructive pulmonary disease. By high-throughput screening of differentially expressed genes in patients with the chronic obstructivepulmonary disease and normal persons, large-sample QPCR verification proves that the content of the DEFA1B gene in the blood of the patients with the chronic obstructive pulmonary disease is higher than the content of the DEFA1B gene in the blood of the normal persons, so that the DEFA1B gene can be used as a biological marker for diagnosing the chronic obstructive pulmonary disease. The researchresult of the invention provides a non-invasive method for early diagnosing the chronic obstructive pulmonary disease clinically, and the method is suitable for being popularized clinically.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a chronic obstructive pulmonary disease diagnostic kit and the use of DEFA1B gene in preparing the chronic obstructive pulmonary disease diagnostic kit. Background technique [0002] Chronic obstructive pulmonary disease (COPD for short) is a common airflow-limited respiratory disease. The airflow limitation usually develops progressively, accompanied by chronic airway inflammation after inhalation of harmful gases or particles. The main clinical manifestations are dyspnea, chronic cough, and cough fatigue. [0003] Worldwide, the prevalence of COPD varies greatly across countries. A survey of people over the age of 40 in five major cities in Latin America, including Mexico City and Montevideo, showed that the prevalence of COPD ranged from 7.8% to 19.7%. Optimistically, a survey data in 2007 showed that the prevalence rate of COPD among people aged 40 and above in 7 regions including ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158
Inventor 刘敏任雁宏
Owner CHINA JAPAN FRIENDSHIP HOSPITAL
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