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Marker for detecting pediatric asthma

A technology of children's asthma and markers, which is applied in the field of markers for children's asthma detection, can solve the problems of unknown wheezing in children, difficulty in bronchial provocation test, lack of diagnostic methods and index gold standards, etc., to achieve rapid detection, The effect of high sensitivity

Active Publication Date: 2013-01-02
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA +1
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  • 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
Diagnosis is mainly based on symptoms and signs, and there is a lack of gold standard for diagnostic methods and indicators

Method used

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  • Marker for detecting pediatric asthma
  • Marker for detecting pediatric asthma
  • Marker for detecting pediatric asthma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1: the selection of children with asthma and contrast

[0018] According to the diagnostic criteria of the guidelines for the diagnosis and prevention of bronchial asthma in children (revised in 2008), in the Respiratory Department of Children's Hospital, clinicians selected children with asthma who met the criteria, aged ≤15 years old, a total of 60 children. A total of 60 healthy children matched in age, sex, and asthma group were selected as the control group.

[0019] The determination of cases depends on the age of the child and selects different criteria:

[0020] 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) other diseases that...

Embodiment 2

[0022] Example 2: Collection of blood samples and detection of biological indicators

[0023] 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.

[0024] Analytical method: Western blot method was used to quantitatively detect the total IgE level in serum.

[0025] 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

[0026] Example 3: Extract RNA

[0027] 3ml of venous blood was extracted from the research subjects, anticoagulated with EDTA to extract RNA. For the extraction method of RNA, please refer to the instruction manual of the ultra-pure total RNA rapid extraction kit produced by Gaining Biotechnology Co., Ltd. After the RNA was extracted from each blood sample, the integrity of the extracted RNA was detected by agarose gel electrophoresis, and its concentration was detected by an ultraviolet spectrophotometer. Calculate the amount of reverse transcription required for 3 μg.

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Abstract

The invention relates to a marker for detecting pediatric asthma, belonging to the field of auxiliary diagnosis of pediatric asthma. The amplified product of semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) or real-time quantitative PCR is adopted as the marker, and is a specific primer aiming at a transient receptor potential vanilloid 2 (TRPV2) gene sequence; an amplification template is total RNA extracted from peripheral blood lymphocyte of a child and is transformed into cDNA by reverse transcription; and the amplified product has the length of 578bp and 95bp. When being used for detecting the TRPV2 gene expression level in the peripheral blood lymphocyte of the child in the asthma group, the marker is higher in sensitivity compared with detection of total IgE.

Description

technical field [0001] The invention belongs to the field of auxiliary diagnosis of children's asthma, and in particular relates to a marker 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. In recent decades, the prevalence of childhood asthma has shown a clear upward trend, and some countries have even more than doubled the prevalence in just 10 years [1] . The results of the national investigation of childhood asthma in my country in 1990 and 2000 showed that the prevalence of childhood asthma increased from 0.91% to 1.50% in 10 years, and the prevalence increased by 64.84%. [2] [0003] Asthma not only seriously endangers the healthy growth of children and has a long-term impact on the quality of life throughout their lives,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 蔡欣徐东群徐东刚付文亮邹民吉朱泽轶
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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