Gene marker RGL4 of sepsis

A sepsis and gene technology, applied in the field of sepsis gene marker RGL4, to achieve the effect of reducing mortality and timely gene diagnosis

Inactive Publication Date: 2017-01-04
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, about a thousand markers are used in sepsis-related research, such as CRP, PCT, activated protein C (APC), high mobility group box B (HMGB1), cytokines, endotoxin, macrophage motility Inhibitor factor (MIF) and other new markers, etc., but there is still a lack of effective and specific markers that can efficiently and easily identify the etiology of inflammation, identify potential viral or bacterial infections, and accurately reflect the efficacy of anti-infection

Method used

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  • Gene marker RGL4 of sepsis
  • Gene marker RGL4 of sepsis
  • Gene marker RGL4 of sepsis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening for gene markers associated with sepsis

[0041] 1. Sample collection

[0042] Blood samples from 10 healthy people and septic patients were collected respectively. All the above samples were obtained with the consent of the ethics committee and the subjects' informed consent.

[0043] 2. RNA sample preparation and quality analysis

[0044] 2.1 Preparation of RNA samples

[0045] (1) Homogenization treatment

[0046] Take blood directly, add 3 times the volume of erythrocyte lysate, mix well, place at room temperature for 10 minutes, and centrifuge at 10,000 rpm for 1 minute. Discard the supernatant thoroughly and collect the white blood cell pellet. Add 1ml Trizol per 100-200μl blood collected leukocyte pellet.

[0047] (2) layered

[0048] a. After adding Trizol to the sample, place it at room temperature for 5 minutes to fully lyse the sample. Centrifuge at 12,000rpm at 4°C for 10min, and take the supernatant;

[0049] b. Add 200 μl of chl...

Embodiment 2

[0071] Example 2 QPCR sequencing to verify the differential expression of the RGL4 gene

[0072] 1. According to the detection results of high-throughput sequencing, the RGL4 gene was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 80 cases of blood from patients with sepsis and 80 cases of blood from healthy people were selected.

[0073] 2. The RNA extraction steps are the same as in Example 1.

[0074] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0075] (1) Take 2 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT), and mix well; 70°C water bath; immediately after 5 min, ice bath for 2-3 min;

[0076](2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), 40 U / μl of RNasin, 200 U / μl of M-MLV, and make up to 25 μl of nuclease-free water;

[0077] (3) After 60 min in water b...

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Abstract

The invention discloses a gene marker RGL4 of sepsis. The expression of the RGL4 gene in sepsis is up-regulated, so that early diagnosis can be provided for a patient with sepsis by detecting the expression level of the RGL4 gene, and early treatment and reduction of the case-fatality rate of the patient with sepsis are expected to be realized.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a gene marker RGL4 of sepsis. Background technique [0002] Sepsis refers to a systemic inflammatory response syndrome due to infection and can lead to severe sepsis (acute organ dysfunction secondary to infection) or septic shock (severe sepsis combined with a hypotensive state irreversible by fluid resuscitation) . Sepsis accounts for about 40% of the total number of patients admitted to the ICU ward, and the mortality rate can be as high as 25%-40%. Early diagnosis and treatment of sepsis are important factors affecting the mortality of patients with sepsis. Sepsis usually has obvious clinical manifestations, but these clinical manifestations are not unique to infection, and non-infectious diseases sometimes manifest as sepsis Toxic features. The use of antibiotics is often based on these non-specific clinical manifestations, which not only affect the diagnosis of sepsis, but also...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/68G01N2333/47
Inventor 杨承刚宋宏涛
Owner BEIJING MEDINTELL BIOMED CO LTD
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