Molecular marker for child respiratory syncytial virus infection and application of molecular marker

A technology of molecular markers and markers, applied in the field of biomedicine, can solve the problem of less research on the role of host factors

Pending Publication Date: 2019-03-22
赵秀侠
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is a certain understanding of how RSV affects host immune defense during infection, but there are few studies on the role of host factors in RSV infection, especially the lack of comprehensive understanding of host factors related to RSV replication at the genome level

Method used

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  • Molecular marker for child respiratory syncytial virus infection and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Screening for Gene Markers Related to Children's Respiratory Syncytial Virus Infection

[0046] 1. Sample collection

[0047] The blood samples of 5 normal children and 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.

[0048] 2. RNA sample preparation and quality analysis

[0049] 2.1 Total RNA was extracted using the RNA extraction kit from Promega. Specific steps are as follows:

[0050] 1) Take 1ml of whole blood collected in a test tube treated with heparin or EDTA, put it into a sterile centrifuge tube, centrifuge at 3000rpm for 5min, and carefully suck off the supernatant from the top of the sample;

[0051] 2) Add 1ml of blood cell lysate, pipette carefully 4-5 times, resuspend the sediment, and centrifuge at 3000rpm for 5min;

[0052] 3) Repeat step 2) twice;

[...

Embodiment 2

[0073] Example 2 QPCR sequencing to verify the differential expression of genes

[0074] 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, blood from 30 children with respiratory syncytial virus infection and 25 normal children were selected.

[0075] 2. RNA extraction steps are the same as in Example 1

[0076] 3. Reverse transcription:

[0077] (1) Reverse transcription reaction:

[0078] Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components to PCR tubes: DEPC water, 5× reverse transcription buffer, 10mM dNTP, 0.1mM DTT, 30μM Oligo dT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1 hour, then centrifuge briefly at 72°C for 10 minutes.

[0079] (2) Primer design

[0080] QPCR amplification primers were designed according to the coding sequences of H2AFJ and RGL4 gen...

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Abstract

The invention discloses a molecular marker for child respiratory syncytial virus infection and application of the molecular marker. According to the invention, up-regulated expression of H2AFJ and RGL4 in child patients with the child respiratory syncytial virus infection is found for the first time; by detecting the expression level of the H2AFJ and the RGL4, whether a subject is suffered from respiratory syncytial virus infection diseases or not can be judged.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a molecular marker of respiratory syncytial virus infection in children and an application thereof. Background technique [0002] Respiratory syncytial virus (Respiratory syncytial virus, RSV) is an important respiratory virus that infects humans. It mainly infects children, the elderly, and people with low immunity. It can cause severe respiratory diseases including pneumonia and bronchitis. One of the leading causes of death. Although some progress has been made in the research of vaccines and therapeutic antibodies against RSV, there are still no commercial vaccines and effective antiviral interventions. The study of viral host factors is helpful to discover new antiviral targets, and is a research hotspot and focus in the current field. [0003] At present, there is a certain understanding of how RSV affects host immune defense during infection, but there are few studies on the role...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/6893G01N2333/47
Inventor 赵秀侠
Owner 赵秀侠
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