Primers and kits for detecting polymorphic sites of asthma susceptible genes ALOX5 (Arachidonate 5-Lipoxygenase), LTA4H (Leukotriene A4Hydrolase) and LTC4S (Leukotriene C4Synthase), and application of primers and kits

A polymorphic site, susceptibility gene technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., can solve the problems of lack of detection kits, few research reports, etc., and achieve short detection time , low price, and the effect of reducing testing costs

Inactive Publication Date: 2018-10-16
浙江梅欧信息科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the need for personalized treatment of clinical bronchial asthma is increasingly prominent, but there are few research reports on the three loci of ALOX5-rs2115819, LTA4H-rs2660845, and LTC4S-rs730012, and the relevant detection kits are even more lacking

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  • Primers and kits for detecting polymorphic sites of asthma susceptible genes ALOX5 (Arachidonate 5-Lipoxygenase), LTA4H (Leukotriene A4Hydrolase) and LTC4S (Leukotriene C4Synthase), and application of primers and kits
  • Primers and kits for detecting polymorphic sites of asthma susceptible genes ALOX5 (Arachidonate 5-Lipoxygenase), LTA4H (Leukotriene A4Hydrolase) and LTC4S (Leukotriene C4Synthase), and application of primers and kits
  • Primers and kits for detecting polymorphic sites of asthma susceptible genes ALOX5 (Arachidonate 5-Lipoxygenase), LTA4H (Leukotriene A4Hydrolase) and LTC4S (Leukotriene C4Synthase), and application of primers and kits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1 Discovery of ALOX5-rs2115819, LTA4H-rs2660845, LTC4S-rs730012 gene variations.

[0079] Gene variation analysis was performed on 3 children with asthma, the detailed steps are as follows:

[0080] 1. Using variation analysis reagents for detection: the reagents include 1) DNA extraction system reagents; 2) DNA amplification system reagents; 3) DNA sequencing system reagents.

[0081] 2. Take 2ml of EDTA anticoagulated peripheral blood samples from suspected pediatric asthma patients and normal control samples, carry out DNA extraction, DNA amplification, and obtain PCR products. The results are shown in agarose electrophoresis ( figure 1 ); PCR product sequencing results ( figure 2 ,3,4).

[0082] Asthma is a polygenic genetic disease. In this study, based on the actual situation of the patients, it was found that when 2 of the 3 patients were treated with Montelukast Sodium Tablets, the therapeutic effect was not obvious. Gene polymorphism analysis found ...

Embodiment 2

[0083] Embodiment 2 Application method and application example of diagnostic kit of the present invention

[0084] 1. Obtain clinical samples and set up experimental and control groups

[0085] 1) Patient A

[0086] Male, 4 years old, from a tertiary hospital. Recurrent cough and asthma for more than 2 years. Physical examination: a small amount of wheezing can be heard in both lungs. Diagnosis considers "bronchial asthma in children".

[0087] Doctor's order:

[0088] 1. Take proper exercise to enhance your physique and avoid catching cold.

[0089] 2. Montelukast sodium tablets (Singulair), 5 mg orally before going to bed.

[0090] 3. Budesonide suspension (Pulmicort Rescue) for inhalation, 1 mg nebulized inhalation twice a day.

[0091] 4. Terbutaline sulfate nebulized liquid (bricanyl), 2.5mg nebulized inhalation three times a day.

[0092] 5. If symptoms worsen, seek emergency treatment at any time.

[0093] Follow-up records: The patient's nebulization was effec...

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Abstract

The invention discloses primers and kits for detecting polymorphic sites of asthma susceptible genes ALOX5 (Arachidonate 5-Lipoxygenase), LTA4H (Leukotriene A4Hydrolase)and LTC4S (Leukotriene C4Synthase), and application of the primers and the kits. Asthma susceptible gene ALOX5, LTA4H and LTC4S reagents comprise the variation analysis kits. The kits comprise (1) a DNA extracting system kit used for extracting peripheral venous blood DNA of a detected individual, (2) a DNA amplification system kit and (3) a DNA sequencing system kit. As for the DNA sequencing system kit, by taking DNA as a template, three primer pairs shown as SEQ ID NO.1-6 are subjected to PCR amplification, and amplified PCR products are sequenced and compared with the gene coded DNA sequence. When the ALOX5-rs2115819 mutant gene type is AG, the therapeutic effect on an asthma patient is not obvious, and when LTA4H-rs2660845 site is A / G, the therapeutic effect on the asthma patient is not obvious. The detecting reagents have the time saving effect and are accurate and low in detection cost.

Description

technical field [0001] The invention relates to the fields of molecular biology and gene detection, in particular to a primer and a kit for detecting polymorphic sites of asthma susceptibility genes ALOX5, LTA4H and LTC4S and the application of the primer and the kit. Background technique [0002] Asthma is a common chronic respiratory disease that seriously endangers human health. It is a chronic airway inflammation characterized by airway hyperresponsiveness, and is involved in a variety of cells and cytokines. The incidence of bronchial asthma is increasing year by year all over the world, and winter and spring in northern my country are the seasons of high incidence. Generally, the mechanism of treating children with asthma or cough variant asthma is similar, and the treatment methods are mainly anti-inflammatory, anti-asthma, and application of hormones. The pathogenesis of bronchial asthma has not yet been clarified, and the current treatment can only achieve the effe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 李智沈哲
Owner 浙江梅欧信息科技有限公司
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