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Biomarkers for the diagnosis of respiratory viral infections in children

A respiratory and children's technology, applied in the field of biomedicine, can solve the problems of lack of pathogenesis, affect the host's antiviral response and virus replication, and achieve the effect of accurate diagnosis

Active Publication Date: 2019-06-04
李然然
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RSV infection can regulate host gene expression, affect host antiviral response and viral replication, but there is still a lack of effective genes to analyze the pathogenesis of respiratory syncytial virus infection in children

Method used

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  • Biomarkers for the diagnosis of respiratory viral infections in children
  • Biomarkers for the diagnosis of respiratory viral infections in children

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Example 1 Screening for Gene Markers Related to Children's Respiratory Syncytial Virus Infection

[0100] 1. Sample collection

[0101] Five normal children's blood and blood samples from children with respiratory syncytial virus infection were collected, and the sample name, number, sampling date, sample processing process, etc. were written. All the above samples were obtained with the consent of the ethics committee.

[0102] 2. Preparation of RNA samples

[0103] RNA samples were extracted using Invitrogen's Blood RNA Extraction Kit. For details, see the instruction manual.

[0104] 3. Quality analysis of RNA samples

[0105] The above extracted RNA was subjected to agarose gel electrophoresis, the concentration and purity of the extracted RNA were detected by Nanodrop2000, the integrity of the RNA was detected by agarose gel electrophoresis, and the RIN value was determined by Agilent2100. The total amount of RNA required for a single library construction is 5ug...

Embodiment 2

[0114] Example 2 QPCR sequencing to verify the differential expression of genes

[0115] 1. According to the detection results of high-throughput sequencing, the differentially expressed genes were verified by large-sample QPCR. According to the sample collection method in Example 1, 45 cases of blood from patients with RSV infection and 18 cases of normal children's blood were selected.

[0116] 2. RNA extraction

[0117] RNA samples were extracted using Invitrogen's Blood RNA Extraction Kit. For details, see the instruction manual.

[0118] 3. Reverse transcription:

[0119] The reverse transcription kit of TAKARA Company was used for operation. Specific steps are as follows:

[0120] (1) Take 2 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT), and mix well. Immediately after 5 minutes in 70°C water bath, ice bath for 2-3 minutes.

[0121] (2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), 40...

Embodiment 3

[0148] ROC analysis of embodiment 3 differentially expressed genes

[0149] Combining the 4 differentially expressed genes screened above, using the ROC curve method and calculating the area under the curve (AUC) to evaluate the diagnostic value of significant influencing factors for respiratory syncytial virus infection in children, AUC and its 95% confidence interval Evaluated by MedCalc, according to different AUC values, it can be divided into no prediction performance (AUC<0.5), low prediction performance (0.5≤AUC≤0.7), moderate prediction performance (0.7≤AUC≤0.9) and high prediction performance (0.9≤ AUC≤1), P<0.05 means the difference is statistically significant.

[0150] The results are shown in Table 1. The area under the curve for the diagnosis of respiratory syncytial virus infection in children by gene combined detection ranged from 0.878 to 0.986.

[0151] Table 1 Area under the curve of differentially expressed genes

[0152] Gene

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Abstract

The invention discloses a biological marker for diagnosing respiratory syncytial virus infection of children. The biological marker can be used for assisting to diagnose the respiratory syncytial virus infection of the children. The invention also discloses a combination of at least two biological markers correlated with the respiratory syncytial virus infection of the children; the respiratory syncytial virus infection of the children can be combined with a bioinformatics method to use; the invention further discloses a product correlated with the biological marker.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a biomarker for diagnosing children's respiratory virus infection. Background technique [0002] Respiratory syncytial virus (Respiratory syncytial virus, RSV) is one of the clinically important pathogens of respiratory tract infection in children in recent years. The virus infects worldwide and has a clear seasonal epidemic trend, with a high incidence in winter and spring. RSV can cause upper respiratory tract infection symptoms (such as colds, etc.), and can also cause lower respiratory tract infection symptoms (such as bronchiolitis and pneumonia, etc.). According to the U.S. Centers for Disease Control (CDC), more than half of children are infected with RSV before the age of 1, and almost all children have at least one RSV infection by the age of 2. 25% to 40% of children have symptoms of bronchiolitis and pneumonia, 0.5% to 2% of children need hospitalization due to RSV infection, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70G01N33/569
CPCC12Q1/701C12Q2600/158G01N33/56983
Inventor 李然然
Owner 李然然
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