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Application of protein S as biological marker in preparation of reagent for diagnosis of asthma and evaluation of prognosis effect

A technology of disease diagnosis and diagnostic markers, applied in the field of biomedicine, to achieve the effect of simple and easy sampling and detection operations

Inactive Publication Date: 2018-09-18
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This protein is a naturally occurring anticoagulant with multiple biological functions, and can participate in physiological reactions such as cell proliferation, apoptosis, blood coagulation, regulation of inflammatory factor release, angiogenesis, and tumorigenesis. Discovered the relationship between Pros1 and asthma

Method used

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  • Application of protein S as biological marker in preparation of reagent for diagnosis of asthma and evaluation of prognosis effect
  • Application of protein S as biological marker in preparation of reagent for diagnosis of asthma and evaluation of prognosis effect
  • Application of protein S as biological marker in preparation of reagent for diagnosis of asthma and evaluation of prognosis effect

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Screening of genes specifically expressed in peripheral blood of asthmatic patients:

[0028] 1. Experimental materials

[0029] 1.1 Peripheral blood cells from asthma patients or healthy people

[0030] Human peripheral blood samples were collected by Guangzhou Women and Children's Medical Center (GWCMC).

[0031] 1.2 Reagents

[0032] Human lymphocyte separation medium Ficol was purchased from GE Healthcare, RNAiso Plus was purchased from TAKARA Company, primers were synthesized by tsingke Company, and SYBR Green Real time PCR Master mix was purchased from Quanshijin Company.

[0033] 1.3 Experimental Instruments

[0034] Quantitative RCP instrument (Stratagene Mx3000P) was purchased from Agilent Company.

[0035] 2. Experimental methods and results

[0036] 2.1 Preparation of cDNA from peripheral blood samples

[0037] (1) Collect peripheral blood from asthmatic patients or healthy people, separate peripheral blood mononucleated cells (PBMCs), resuspend in 1 ml...

Embodiment 2

[0048] Detection of Pros1 protein level in peripheral blood of asthmatic patients:

[0049] 1. Experimental materials

[0050] 1.1 Peripheral blood serum of asthma patients or healthy people

[0051] Human peripheral blood samples were collected by Guangzhou Women and Children's Medical Center (GWCMC).

[0052] 1.2 Reagents

[0053] Protein S (PROS) detection kit.

[0054] 1.3 Experimental Instruments

[0055] The multi-label detection system was purchased from EnSpire Company.

[0056] 2. Experimental methods and results

[0057] 2.1 Collection and freezing of peripheral blood samples

[0058] Peripheral blood serum was collected from asthma patients (including mild and severe asthma) or healthy people, and stored at -80°C.

[0059] 2.2 Quantitative determination of Pros1 in human serum by double antibody sandwich ELISA

[0060] Coat the Pros1 antibody in a 96-well plate to make a solid phase carrier, add standards or samples to the microwells, where Pros1 binds to the ...

Embodiment 3

[0072] The relationship between the protein level of Pros1 and the percentage of eosinophils in alveolar lavage fluid of mouse asthma model

[0073] 1. Experimental materials

[0074] 1.1 Mouse asthma model

[0075] Wild-type C57BL / 6 mice aged 6 to 8 weeks were purchased from Beijing Weitong Lihua and raised in an SPF clean-grade barrier environment in the animal room of Wuhan Institute of Virology, Chinese Academy of Sciences. On days 0, 7, and 14, intraperitoneal injection of 50 micrograms of chicken ovalbumin emulsified with aluminum adjuvant was performed for sensitization, and intranasal immunization with 50 micrograms of chicken ovalbumin was performed continuously on days 21, 22, and 23, and sacrificed 1 day later For mice, alveolar lavage fluid (BAL) was collected.

[0076] 1.2 Reagents

[0077] Mouse vitamin K-dependent Pros1 (PROS1) ELISA kit was purchased from Wuhan Huamei Bioengineering Co., Ltd. Antibody Silgec-F-APC, CD11c-PE, Fixable Viability Dye 506 was p...

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Abstract

The invention relates to the field of biological medicines and discloses application of protein S (Pros1) as a biological marker in preparation of a reagent for diagnosis of asthma and evaluation of the prognosis effect. The inventors find that the RNA level of Pros1 in peripheral blood of patients suffering from asthma is obviously higher than that of healthy people; further, the ELISA detectionis further performed on Pros1 in peripheral blood of patients suffering from mild and severe asthma and healthy people and finds that the protein level of Pros1 in peripheral blood of patients suffering from severe asthma is obviously higher than that of healthy people; and furthermore, for patients suffering from asthma, the protein level of Pros1 in peripheral blood of patients in the acute stage is higher than that of patients in the remission stage, so that the protein S can serve as the biological marker for diagnosis of asthma and evaluation of the prognosis effect.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of protein S (Protein S, Pros1) as a biomarker in the preparation of reagents for the diagnosis and prognosis of asthma. Background technique [0002] Asthma is one of the most common chronic immune diseases, affecting people of all ages. It is known as one of the four major killers that seriously endanger the quality of human life, second only to AIDS, tumors and cardiovascular and cerebrovascular diseases. About 300 million people worldwide suffer from asthma, and at least 250,000 people die from asthma-related diseases every year. In recent years, with air pollution and environmental deterioration, the incidence of asthma has increased year by year. The current situation of asthma in my country is that the prevalence rate is still relatively low, but the prevalence rate is growing very fast. An epidemiological survey conducted by the China Asthma Alliance in 2013 c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/106C12Q2600/118G01N33/68G01N2800/122
Inventor 魏滨邓力程梦兰
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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