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Application of intestinal flora in the diagnosis of repeated respiratory tract infections in children

A technology for the respiratory tract and children, applied in the field of biomedicine, can solve the problem of unclear pathogenesis, and achieve the effect of strong sensitivity and high specificity

Inactive Publication Date: 2019-10-08
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the pathogenesis of RRT is still unclear

Method used

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  • Application of intestinal flora in the diagnosis of repeated respiratory tract infections in children
  • Application of intestinal flora in the diagnosis of repeated respiratory tract infections in children
  • Application of intestinal flora in the diagnosis of repeated respiratory tract infections in children

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Screening of intestinal flora associated with children's recurrent respiratory tract infection

[0034] 1. Collection of samples

[0035] The feces of 20 healthy children and 20 cases of children with recurrent respiratory tract infection were collected, and the specimens were frozen and stored in a -80°C refrigerator. The detailed clinical information of the samples was recorded, including gender, age, frequency of upper and lower respiratory tract infections, etc. This study was reviewed and approved by the ethics committee, and informed consent was obtained from the patients and their guardians.

[0036] 2. 16S rRNA sequencing

[0037] 2.1 Extraction of DNA

[0038] Bacterial DNA was extracted from stool samples using a stool genomic DNA extraction kit, and the operating steps were performed according to the instructions.

[0039] 2.2 DNA sample purity and concentration determination

[0040] Genomic DNA was detected by 1% agarose gel electrophoresis. ...

Embodiment 2

[0058] Example 2 Fluorescence quantitative PCR verification related flora

[0059] 1. Perform large-scale QPCR verification on the above-mentioned flora, and collect 60 cases of stool samples from healthy children and children with recurrent respiratory tract infection according to the sample collection method in Example 1.

[0060] 2. Extraction and quantification of fecal bacterial DNA

[0061] Bacterial DNA was extracted from stool samples using a stool genomic DNA extraction kit, and the operation steps were carried out according to the instructions;

[0062] Genomic DNA was detected by 1% agarose gel electrophoresis to determine the total DNA concentration and purity of the extracted fecal bacteria, and the mass concentration of DNA in all samples was unified to 100 mg / L.

[0063] 3. Real-time fluorescent quantitative PCR

[0064] 1) Primer design and synthesis

[0065] According to the V3-V4 region sequence of bacterial 16SrRNA gene, specific primers were designed for...

Embodiment 3

[0079] The preparation of embodiment 3 children's recurrent respiratory tract infection diagnostic kit

[0080]According to the correlation between Fusicatenibacter, Roseburia and children's recurrent respiratory tract infection disease, children's recurrent respiratory tract infection disease can be diagnosed by detecting the abundance of Fusicatenibacter, Roseburia in the sample, and accordingly the present invention provides a kind of diagnosis based on the detection of Fusicatenibacter, Roseburia abundance Kit for recurrent respiratory infections in children. The kit components are as follows: DNA extraction reagents, primer pairs for specific detection of Fusicatenibacter and Roseburia (Roseburia) 16SrRNA, reaction buffer, deoxynucleotide triphosphate bases (dNTPs), Taq-polymerase reverse Recording enzyme, DNase, RNAse inhibitor, DEPC-water, sterile water, SYBR Green fluorescent dye.

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Abstract

The invention discloses application of an intestinal flora to diagnosis of recurrent respiratory tract infection of children. Through 16SrRNA sequencing on a child patient suffering from the recurrentrespiratory tract infection, existence of the flora with abundance difference in the patient is found for the first time; further analysis on the differential flora finds that during application to the diagnosis of the disease, the flora has relatively high sensitivity and specificity, which prompts that early accurate diagnosis of the recurrent respiratory tract infection of the children can beachieved through detection on the abundance of the flora.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of intestinal flora in children's recurrent respiratory tract infection. Background technique [0002] Recurrent respiratory tract infection (RRTI) is an important clinical disease, especially in pediatrics, which is common and frequent. The clinical features of RRTI are recurrent upper and lower respiratory tract infections, and the duration of infection in children is longer than that of normal children. Once the treatment is not timely, it will spread rapidly and aggravate the condition. Some children have attacks several times a month, and some children have a course of several years. The disease is recurrent, protracted and difficult to heal, which can lead to various complications. [0003] In recent years, its incidence rate has been increasing year by year, and RRTI accounts for about 10-20% of the cases of respiratory tract infection. According to the World Hea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/04C40B40/06
Inventor 李蕾
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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