CN116223818A - Application of DR3 protein detection reagent in preparation of reagent kit for screening AECOPD and reagent kit for screening AECOPD - Google Patents
Application of DR3 protein detection reagent in preparation of reagent kit for screening AECOPD and reagent kit for screening AECOPD Download PDFInfo
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- CN116223818A CN116223818A CN202310516991.8A CN202310516991A CN116223818A CN 116223818 A CN116223818 A CN 116223818A CN 202310516991 A CN202310516991 A CN 202310516991A CN 116223818 A CN116223818 A CN 116223818A
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6884—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from lung
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Abstract
The invention provides an application of a DR3 protein detection reagent in preparing a kit for screening AECOPD and the kit for screening AECOPD, and belongs to the field of in-vitro diagnostic reagents. The invention discovers that the expression level of DR3 protein in serum of AECOPD patients is obviously higher than that of common COPD patients and healthy people for the first time. The reagent for detecting the expression level of the DR3 protein is used for preparing the AECOPD screening kit, so that the effective screening of AECOPD can be realized, and the reagent has good clinical application prospect.
Description
Technical Field
The invention belongs to the field of in-vitro diagnostic reagents, and particularly relates to application of a DR3 protein detection reagent in preparing a kit for screening AECOPD and the kit for screening AECOPD.
Background
Chronic obstructive pulmonary disease (Chronic obstructive pulmonary disease, COPD) is a common pulmonary disease characterized by persistent respiratory symptoms and airflow limitation that can further progress to pulmonary heart disease and respiratory failure. COPD is associated with abnormal inflammatory reactions of noxious gases and noxious particles, has high disability and mortality rates, has a global morbidity of 9% -10% over 40 years old, and is one of the diseases with the highest morbidity and mortality rates worldwide.
The acute exacerbation phase of chronic obstructive pulmonary disease (Acute exacerbation of pulmonary disease, AECOPD) is an important event in the clinical course of COPD, and COPD patients experience about 0.5-3.5 acute exacerbations each year, and frequent acute exacerbations of chronic obstructive pulmonary disease can cause exacerbation of pulmonary function, so AECOPD is an important factor in the death of COPD patients. AECOPD in the revised guidelines for diagnosis and treatment of chronic obstructive pulmonary disease in 2007 in China refers to the manifestation of significant exacerbation of inflammation such as fever, which is caused by persistent exacerbation of the patient beyond the daily condition and need to change the conventional medication of basic COPD, usually in the course of the disease, in which the patient suffers from exacerbation of cough, expectoration, shortness of breath and/or wheezing in a short period, and increased sputum volume, which is purulent or mucopurulent. The treatment method of AECOPD mainly delays disease progression and relieves symptoms, so early diagnosis and early treatment are important for AECOPD. However, the definition and diagnosis of AECOPD are all descriptions of clinical symptoms at present, and lack of quantitative indexes is easy to cause missed diagnosis and misdiagnosis.
With the continuous development of molecular biology, biomarkers as a quantitative index for disease diagnosis have become a research hotspot. In recent years, various biomarkers have been found to be involved in the development of COPD, such as VEGF, IL-6, IL-8, TNF- α, and the like. But lack of biomarkers of excellent predictive efficacy for AECOPD with high specificity and sensitivity. Death receptor 3 (DR 3) is a member of the Tumor Necrosis Factor Superfamily (TNFSF) and forms a receptor-ligand complex with tumor necrosis factor ligand-1A (TL 1A), which regulates immunity, inflammation, proliferation and death of cells, and is involved in the progression of various diseases. There is currently no study that demonstrates a clear relationship between DR3 and AECOPD diagnosis.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides an application of a DR3 protein detection reagent in preparing a kit for screening AECOPD and a kit for screening AECOPD.
The invention provides an application of a reagent for detecting DR3 protein in preparing a screening kit for acute exacerbation of chronic obstructive pulmonary disease.
Further, the reagent for detecting DR3 protein is enzyme-linked immunoassay reagent.
Further, the reagent for detecting the DR3 protein is a western blot reagent.
Further, the reagent for detecting DR3 protein is a reagent for a protein chip detection method.
Further, the reagent for detecting DR3 protein is a reagent for detecting DR3 protein in human serum.
The invention also provides a screening kit for acute exacerbation of chronic obstructive pulmonary disease, which comprises a reagent for detecting DR3 protein.
Further, the reagent for detecting DR3 protein is enzyme-linked immunoassay reagent.
Further, the reagent for detecting the DR3 protein is a western blot reagent.
Further, the reagent for detecting DR3 protein is a reagent for a protein chip detection method.
Further, the reagent for detecting DR3 protein is a reagent for detecting DR3 protein in human serum.
The key point of the invention is to determine whether the expression level of DR3 protein in serum of a person to be detected is obviously related to the condition of AECOPD, and compared with common COPD patients and healthy people, the risk of AECOPD is high when the expression level of DR3 protein is high, so that whether the person to be detected has AECOPD can be screened by detecting the expression level of DR3 protein in serum of the person to be detected. As means for specifically detecting the expression level of DR3 protein in serum, various means disclosed in the prior art can be employed, not only the methods used in the examples of the present invention. Any method capable of detecting DR3 protein expression levels in serum may be used in the present invention.
Compared with the prior art, the invention has the beneficial effects that:
the research of the invention finds that the expression level of DR3 protein is specifically related to AECOPD patients, and the expression level of DR3 protein in serum of AECOPD patients is obviously higher than that of common COPD patients and healthy people. Thus, AECOPD can be screened by detecting DR3 protein expression levels in patient serum. If the DR3 protein expression level is high (relative to healthy people and COPD patients), the risk of AECOPD is high, and if the DR3 protein expression level is low, the risk of AECOPD is low. The expression level of DR3 protein in serum can be used for diagnosis or auxiliary diagnosis of clinical AECOPD, provides effective basis for relevant therapeutic measures or decisions of patients, and has good clinical application prospect.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 shows the expression levels of DR3 protein in serum of COPD patients, AECOPD patients and healthy volunteers.
FIG. 2 is an ROC analysis of the expression level of DR3 protein in serum of COPD patients and AECOPD patients.
FIG. 3 is an ROC analysis of the expression level of DR3 protein in serum of healthy volunteers and AECOPD patients.
Detailed Description
The materials and equipment used in the embodiments of the present invention are all known products and are obtained by purchasing commercially available products.
EXAMPLE 1 relation of DR3 in serum to AECOPD
1. Patient population
Hua Xiyi patients with stable COPD in the outpatient setting 112, 109 patients with Acute Exacerbations (AECOPD) in the hospitalization, and 69 healthy volunteers. Stable COPD patients and healthy volunteers were tested for standard lung function using standard methods adopted by the american society of thoracic guidelines. Diagnosis of chronic obstructive pulmonary disease is based on global chronic obstructive pulmonary disease initiative (GOLD) criteria: (a) After use of the bronchodilator beta/FVC < - > agonist (200 mg salbutamol), (b) Forced Vital Capacity (FVC) or FEV1< 12% or 200 mL increase.
2. Sample detection
The subject's elbow vein was given fasting venous blood, centrifuged directly at 3000rpm for 10 minutes at 4℃and the supernatant was taken. Serum was then saved to analysis at-80 ℃ and DR3 protein expression levels in serum were detected. The detection was performed on a Bio-Plex 200 detection platform (Bio-Rad, california, USA) using a Human Luminex cube detection Assay (LXSAHM) according to manufacturer's instructions, using reagents such as anti-Human DR3 antibody coated magnetic beads, biotin-labeled secondary antibodies, chromogenic substrates, etc., all of which were commercially available. The DR3 detection limit was 58.6pg/mL. The operator taking the measurement is unaware of the clinical information of the subject.
3. Results data
The expression levels of DR3 protein in serum of COPD patients, AECOPD patients and healthy volunteers are shown in table 1 and fig. 1. The expression level data of 3 sets of DR3 proteins were compared and the results are shown in table 2. The ROC analysis results for COPD patients and AECOPD patients are shown in fig. 2, table 3. The ROC analysis results for healthy volunteers and AECOPD patients are shown in fig. 3, table 4.
TABLE 1 expression levels of DR3 protein in serum of COPD, AECOPD patients and healthy volunteers
TABLE 2 comparison of DR3 protein expression levels in serum of COPD, AECOPD patients and healthy volunteers
TABLE 3 ROC Curve analysis results for COPD patients and AECOPD patients
TABLE 4 ROC analysis results for healthy volunteers and AECOPD patients
From the above data, it can be seen that: the level of DR3 protein expression in serum was significantly increased in AECOPD patients compared to COPD patients and healthy volunteers, both statistically different. ROC analysis results for COPD patients and AECOPD patients: AUC area was 0.829, optimal threshold was 117.5950, specificity was 60.7% and sensitivity was 95.4%. ROC analysis results in healthy volunteers and AECOPD patients: AUC area was 0.770, optimal threshold was 240.5700, specificity was 97.1% and sensitivity was 60.6%.
The results show that the DR3 expression level in serum is less different in COPD patients than in healthy persons, whereas the DR3 expression level in serum is significantly different in AECOPD patients than in COPD patients. Therefore, the DR3 protein can specifically distinguish AECOPD patients from healthy people and COPD patients, and the specificity and sensitivity of screening AECOPD patients by taking the DR3 protein as a marker are good. The aim of screening AECOPD can be achieved by detecting the expression level of DR3 protein in serum.
In summary, the present study found that DR3 protein expression levels are specifically associated with AECOPD patients, and that DR3 protein expression levels in serum of AECOPD patients are significantly higher than in normal COPD patients and healthy people. Thus, AECOPD can be screened by detecting DR3 protein expression levels in patient serum. If the DR3 protein expression level is high (relative to healthy people and COPD patients), the risk of AECOPD is high, and if the DR3 protein expression level is low, the risk of AECOPD is low. The expression level of DR3 protein in serum can be used for diagnosis or auxiliary diagnosis of clinical AECOPD, provides effective basis for relevant therapeutic measures or decisions of patients, and has good clinical application prospect.
Claims (10)
1. The reagent for detecting DR3 protein is used in preparing reagent kit for screening acute exacerbation period of chronic obstructive pulmonary disease.
2. Use according to claim 1, characterized in that: the reagent for detecting DR3 protein is enzyme-linked immunoassay reagent.
3. Use according to claim 1, characterized in that: the reagent for detecting the DR3 protein is a western blot reagent.
4. Use according to claim 1, characterized in that: the reagent for detecting DR3 protein is a reagent for a protein chip detection method.
5. The use according to any one of claims 1 to 4, characterized in that: the reagent for detecting the DR3 protein is a reagent for detecting the DR3 protein in human serum.
6. A screening kit for acute exacerbation of chronic obstructive pulmonary disease, which is characterized in that: it includes reagents for detecting DR3 protein.
7. The kit of claim 6, wherein: the reagent for detecting DR3 protein is enzyme-linked immunoassay reagent.
8. The kit of claim 6, wherein: the reagent for detecting the DR3 protein is a western blot reagent.
9. The kit of claim 6, wherein: the reagent for detecting DR3 protein is a reagent for a protein chip detection method.
10. The kit according to any one of claims 6 to 9, wherein: the reagent for detecting the DR3 protein is a reagent for detecting the DR3 protein in human serum.
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