Reagent and reagent kit for detecting children asthma based on GLCCI1 gene rs37972 site and application

Abstract

The invention discloses a reagent and reagent kit for detecting children asthma based on a GLCCI1 gene rs37972 site and an application. The reagent comprises a PCR reaction system, an SAP reaction system and an iPLEX extension system. The reagent kit comprises the reagent. The reagent is applied to preparation of the reagent kit for predicting the risk that children suffer from the asthma through the steps of performing PCR amplification, performing an SAP reaction, performing iPLEX extension, and performing gene sequencing, wherein if a genotype at the GLCCI1 gene rs37972 site is TT or CT, after ICS treatment, MMEF improvement is small. The reagent disclosed by the invention can be used for performing early genetics prediction on the generation of the asthma.

Description

technical field

[0001] The invention relates to the technical field of genetic engineering, in particular to a reagent, kit and application for detecting childhood asthma based on the rs37972 site of the GLCCI1 gene. Background technique

[0002] Bronchial asthma (bronchial asthma, referred to as asthma) is a heterogeneous disease characterized by chronic airway inflammation. Reversible expiratory airflow limitation. There are about 358 million people with asthma in the world, among which 30 million people have asthma in China. According to the survey data, the global average incidence rate of asthma in adults is 4.3%; while the incidence rate of children's asthma is 11.6% in children aged 6-7, and 13.7% in children aged 13-14, and there is a large difference between countries, 6 The incidence of asthma among children aged 7 to 7 was as high as 37.6% (Costa Rica), and among children aged 13 to 14 was as high as 31.2% (Isle of Man). In my country, the survey data in 2010 s...

Examples

Embodiment 1

[0041] A detection reagent for detecting asthma in children, comprising: PCR reaction system, SAP reaction system and iPLEX extension system.

[0042] The total volume of the PCR reaction system is 5 μL, specifically including 1.625mM MgCl 2 , 500 μM dNTP, 50 μM upstream primer, 50 μM downstream primer, 0.5U / rxn Taq enzyme, 1.25x PCR Buffer and deionized water.

[0043] Wherein the DNA sequence of the upstream primer is shown in SEQ ID NO.1, specifically:

[0044] ACGTTGGATGCACGAGCAGACCAATTTGAC.

[0045] The DNA sequence of the downstream primer is shown in SEQ ID NO.2, specifically:

[0046] ACGTTGGATGCGGGAGCAATTAATGTAAGG.

[0047] The total volume of the SAP reaction system is 7 μl, and its components include: 1U / μl shrimp alkaline phosphatase, 10x SAP Buffer and deionized water.

[0048] The total volume of the iPLEX extension system is 9μl, and its components include: 0.222x iPLEX Buffer, 1x iPLEX Termination mix, 1x iPLEX Enzyme, Primer Mix (7μM: 14μM).

Embodiment 2

[0050] A kit for detecting asthma in children, comprising the reagent of Example 1.

Embodiment 3

[0052] An application of the reagent of Example 1 in the preparation of a kit for predicting the risk of children suffering from asthma, the application method comprising the following steps:

[0053] (1) Selection of inspection objects:

[0054] A total of 263 Han children with asthma diagnosed in the Pediatric Outpatient Clinic of Xiangya Hospital, Central South University were recruited, with an average age of 8.18 (±2.73) years old. At the same time, 150 healthy control children with an average age of 8.20 (±2.28) years old were recruited from the Department of Pediatrics, Xiangya Hospital, Central South University. At the same time, the above-mentioned children were divided into a group older than and equal to 6 years old (group A) and a group younger than 6 years old (group B) according to their age.

[0055] Inclusion criteria for asthma group:

[0056] 1. Meet the diagnostic criteria for asthma in the "Guidelines for the Diagnosis and Prevention of Bronchial Asthma i...

technical field

[0001] The invention relates to the technical field of genetic engineering, in particular to a reagent, kit and application for detecting childhood asthma based on the rs37972 site of the GLCCI1 gene. Background technique

[0002] Bronchial asthma (bronchial asthma, referred to as asthma) is a heterogeneous disease characterized by chronic airway inflammation. Reversible expiratory airflow limitation. There are about 358 million people with asthma in the world, among which 30 million people have asthma in China. According to the survey data, the global average incidence rate of asthma in adults is 4.3%; while the incidence rate of children's asthma is 11.6% in children aged 6-7, and 13.7% in children aged 13-14, and there is a large difference between countries, 6 The incidence of asthma among children aged 7 to 7 was as high as 37.6% (Costa Rica), and among children aged 13 to 14 was as high as 31.2% (Isle of Man). In my country, the survey data in 2010 s...

Examples

Embodiment 1

[0041] A detection reagent for detecting asthma in children, comprising: PCR reaction system, SAP reaction system and iPLEX extension system.

[0042] The total volume of the PCR reaction system is 5 μL, specifically including 1.625mM MgCl 2 , 500 μM dNTP, 50 μM upstream primer, 50 μM downstream primer, 0.5U / rxn Taq enzyme, 1.25x PCR Buffer and deionized water.

[0043] Wherein the DNA sequence of the upstream primer is shown in SEQ ID NO.1, specifically:

[0044] ACGTTGGATGCACGAGCAGACCAATTTGAC.

[0045] The DNA sequence of the downstream primer is shown in SEQ ID NO.2, specifically:

[0046] ACGTTGGATGCGGGAGCAATTAATGTAAGG.

[0047] The total volume of the SAP reaction system is 7 μl, and its components include: 1U / μl shrimp alkaline phosphatase, 10x SAP Buffer and deionized water.

[0048] The total volume of the iPLEX extension system is 9μl, and its components include: 0.222x iPLEX Buffer, 1x iPLEX Termination mix, 1x iPLEX Enzyme, Primer Mix (7μM: 14μM).

Embodiment 2

[0050] A kit for detecting asthma in children, comprising the reagent of Example 1.

Embodiment 3

[0052] An application of the reagent of Example 1 in the preparation of a kit for predicting the risk of children suffering from asthma, the application method comprising the following steps:

[0053] (1) Selection of inspection objects:

[0054] A total of 263 Han children with asthma diagnosed in the Pediatric Outpatient Clinic of Xiangya Hospital, Central South University were recruited, with an average age of 8.18 (±2.73) years old. At the same time, 150 healthy control children with an average age of 8.20 (±2.28) years old were recruited from the Department of Pediatrics, Xiangya Hospital, Central South University. At the same time, the above-mentioned children were divided into a group older than and equal to 6 years old (group A) and a group younger than 6 years old (group B) according to their age.

[0055] Inclusion criteria for asthma group:

[0056] 1. Meet the diagnostic criteria for asthma in the "Guidelines for the Diagnosis and Prevention of Bronchial Asthma i...

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