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Marker and primer for detecting pediatric asthma

A childhood asthma and marker technology, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc., can solve problems such as lack of correlation, difficulty in bronchial provocation test, and difficulty in diagnosing asthma in children , to achieve the effect of high sensitivity

Active Publication Date: 2013-11-20
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The diagnosis of asthma in children mainly has the following difficulties: 1. The diagnosis of asthma in children needs to exclude other diseases, but the types of diseases to be excluded are quite large, which requires rich work experience and prudent work attitude of clinicians
2. It is difficult to carry out pulmonary function test and bronchial provocation test for children under 5 years old, which increases the difficulty of diagnosing asthma in children
3. It is difficult to understand recurrent wheezing. For example, the correct grasp of the child's previous wheezing times should be based on medical records rather than parents' chief complaints. However, many parents in my country, especially those from rural areas, often lose medical records due to improper storage. Therefore, the child's previous wheezing status is unknown
The only study that found the GATA3 polymorphism to be associated with asthma patients with high levels of IgE in the Finnish population [8] , while polymorphisms in the GATA3 gene were not associated with asthmatic children in Germany [9]

Method used

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  • Marker and primer for detecting pediatric asthma
  • Marker and primer for detecting pediatric asthma
  • Marker and primer for detecting pediatric asthma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: the selection of children with asthma and contrast

[0017] According to the diagnostic criteria of the Guidelines for the Diagnosis and Prevention of Bronchial Asthma in Children (revised in 2008), clinicians in the Respiratory Department of Children's Hospital and the Capital Institute of Pediatrics selected 230 children with asthma who met the criteria, aged ≤15 years old. A total of 205 healthy children matched in age, sex, and asthma group were selected as the control group.

[0018] The determination of cases depends on the age of the child and selects different criteria:

[0019] Diagnostic criteria for asthma in infants and young children: (1) age < 3 years old, wheezing attack ≥ 3 times; (2) wheezing sound heard in both lungs and expiratory phase during the attack, and the expiratory phase is prolonged; (3) having an atopic constitution, Such as allergic eczema, allergic rhinitis, etc.; (4) parents have a history of asthma or other allergies; (5)...

Embodiment 2

[0021] Example 2: Collection of blood samples and detection of biological indicators

[0022] The venous blood of the research subjects was drawn to detect the total IgE in the serum, and this part of the detection work was assisted by the hospital.

[0023] Analytical method: Western blot method was used to quantitatively detect the total IgE level in serum.

[0024] Specific operation method: extract 1ml of venous blood from the patient, separate the serum, take 250 μL and incubate with the nitrocellulose membrane adsorbed with specific allergens in the reaction tank at room temperature for 45 minutes, add biotin-labeled anti-human IgE antibody after washing, and incubate at room temperature After 45 minutes, the unbound secondary antibody was eluted, and streptavidin conjugated with alkaline phosphatase was added, and then incubated at room temperature for 20 minutes, the unbound enzyme was washed away, and the substrate was added to develop color. All incubations were don...

Embodiment 3

[0025] Embodiment 3: blood sample collection and storage

[0026] Draw 1ml of venous blood into an EDTA anticoagulant tube, mix it upside down to prevent the blood from coagulating into clots. The blood was transferred to a 1.5ml centrifuge tube and centrifuged at 2000rpm for 10 minutes, leaving a layer of plasma for DNA extraction. If the genomic DNA cannot be extracted on the same day, the blood sample extracted within one week can be stored in the refrigerator at 4°C, and the blood sample extracted within one month should be stored in the refrigerator at -20°C, and the extraction time should be longer Blood samples should be stored in a -70°C refrigerator.

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Abstract

The invention discloses a marker and a primer for detecting pediatric asthma, belonging to the technical field of auxiliary diagnosis of pediatric asthma. According to the invention, the genotype of an SNP (single nucleotide polymorphisms) 290G / A site of a GATA3 gene 5' promoter region can be analyzed by adopting a PCR (Polymerase Chain Reaction) method through extracting the genomic DNA from the blood of children in an asthma group, and the relation between the 290G / A of the GATA3 gene and the Han-nationality pediatric asthma in Beijing of China is evaluated. The association research result indicates that the SNP290G / A of the GATA3 gene is associative to the pediatric asthma in Beijing area, is a high-risk factor of the pediatric asthma, and is higher in sensibility compared with the total IgE (immunoglobulin E) detected.

Description

technical field [0001] The invention belongs to the technical field of auxiliary diagnosis of children's asthma, and in particular relates to a marker and a primer for detecting children's asthma. Background technique [0002] Bronchial asthma is the most common chronic lung disease that threatens public health in the world. The occurrence of asthma can affect people of all ages. It can start in infants and young children, and it is more common in children. The 2009 Global Initiative for Asthma (GINA) pointed out that the incidence of asthma in all age groups has increased significantly in the past 20 years, especially in children. In the 1990s, a sample survey on the prevalence of asthma among children aged 0-14 in my country showed that the prevalence of asthma among children in my country was 0.11%-2.03%. [1] , and the survey in 2000 showed that the prevalence of asthma in children in my country was 0.25% to 4.63%. Therefore, the incidence of asthma in children in my cou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113C12N15/11
Inventor 蔡欣徐东刚徐东群王金凤邹民吉付文亮
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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