Molecular diagnosis marker for pyemia

A sepsis, primer pair technology, applied in the field of molecular diagnostic markers of sepsis, can solve the problem of unsatisfactory sensitivity and specificity, cannot be used as a potential target for sepsis treatment, cannot be used for sepsis diagnosis And other issues

Inactive Publication Date: 2016-11-16
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the sensitivity and specificity of these currently used markers are not ideal, and the scoring system can be used to evaluate the prognosis and severity of the condition of patients with sepsis, but it canno...

Method used

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  • Molecular diagnosis marker for pyemia
  • Molecular diagnosis marker for pyemia
  • Molecular diagnosis marker for pyemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0041] 1. Sample collection

[0042] 10 blood samples from normal people and septic patients were collected, and all the above samples were obtained with the consent of the ethics committee.

[0043] 2. RNA sample preparation and quality analysis

[0044] 2.1 Preparation of RNA samples

[0045] Total RNA was extracted using the RNA extraction kit from Promega. Specific steps are as follows:

[0046] 1) Take 1ml of whole blood collected in a test tube treated with heparin or EDTA, and put it into a sterile centrifuge tube;

[0047] 2) Collect blood cells: centrifuge at 3000rpm for 5min, carefully suck off the supernatant from the top of the sample;

[0048] 3) Add 1ml of blood cell lysate, pipette carefully 4-5 times, and resuspend the sediment;

[0049] 4) Centrifuge at 3000rpm for 5min;

[0050] 5) Repeat steps 3) and 4) twice (three times in total);

[0051] 6) Avoid the cell sediment, carefully suc...

Embodiment 2

[0076] Example 2 QPCR sequencing to verify the differential expression of CYB561A3 gene

[0077] 1. According to the detection results of high-throughput sequencing, the CYB561A3 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 normal blood were selected.

[0078] 2. The RNA extraction steps are the same as in Example 1.

[0079] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0080] (1) Take 2 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT) and mix well; 70°C water bath; 5 min and then immediately ice bath for 2-3 min;

[0081] (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;

[0082] (3) After 42°C water bath for 60 m...

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Abstract

The invention discloses a molecular diagnosis marker for pyemia. Specifically, the diagnosis marker is CYB561A3, and the expression level of CYB561A3 in pyemia patients is reduced. By the adoption of the pyemia diagnosis product, early diagnosis of pyemia is made possible, high mortality caused by diagnosis delay is reduced, and medical resources are saved.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a sepsis molecular diagnostic marker, specifically the molecular marker is CYB561A3. Background technique [0002] Sepsis is a medical condition characterized by a severe systemic inflammatory response caused by infection. It is more common in severe trauma, burns, various shocks, and after major surgery. Further development may lead to septic shock and multiple organ dysfunction (MODS), sepsis is currently the leading cause of death in intensive care unit (ICU) patients. About 28.3% to 41% of sepsis patients died due to multiple organ failure. The relevant pathological mechanisms of sepsis are considered to be intricate, and immune promotion and suppression exist contradictoryly in sepsis, and they are synchronized with the development of the disease, showing corresponding changes. [0003] After the invasion of pathogenic microorganisms in the body, there will be a cascade release of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/569
CPCC12Q1/6883C12Q2600/158G01N33/569
Inventor 李婷董东
Owner BEIJING MEDINTELL BIOMED CO LTD
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