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Molecular marker tcons_00016233, kit and application

A technology of molecular markers and kits, which is applied in the field of molecular markers for early diagnosis and prediction of sepsis complicated with acute kidney injury, and can solve problems such as inaccurate reflection of renal function

Active Publication Date: 2018-06-01
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the increase of serum creatinine often lags behind the deterioration of renal function, and cannot accurately reflect the change of renal function

Method used

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  • Molecular marker tcons_00016233, kit and application
  • Molecular marker tcons_00016233, kit and application
  • Molecular marker tcons_00016233, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Screening for molecular markers associated with sepsis complicated by acute kidney injury

[0021] 1. Sample collection

[0022] Blood samples were collected from healthy people, patients with sepsis without acute kidney injury, and patients with sepsis complicated with acute kidney injury.

[0023] 2. RNA sample preparation and quality analysis

[0024] Use the Trizol of Kangwei Century Company to extract total RNA, and the specific operation steps are as follows:

[0025] 1) Take 2ml of whole blood collected in a test tube treated with sodium citrate, and put it into an enzyme-free centrifuge tube;

[0026] 2) Collect plasma: Centrifuge at 3000rpm for 10min, carefully suck the supernatant (plasma) from the top of the sample and put it into another enzyme-free centrifuge tube;

[0027] 3) Take 250 μL of plasma liquid, transfer it to a 1.5ml centrifuge tube, add 750 μL of TRIzol reagent, shake the tube body vigorously by hand until mixed.

[0028] 4) Afte...

Embodiment 2

[0043] Embodiment 2: RT-PCR verification differential expression

[0044]

[0045]

[0046] 1. According to the detection results of high-throughput sequencing, select for RT-PCR verification. According to the sample collection method in Example 1, blood samples from healthy people, sepsis complicated with acute kidney injury, and sepsis without acute kidney injury were selected.

[0047] 2. RNA extraction steps are the same as in Example 1

[0048] 3. Reverse transcription: use the reverse transcription kit from ThermoFish Company to operate, and the specific steps are as follows:

[0049] 1) Add the reagents used, store on ice, and conduct related experiments according to the table below.

[0050] 2) Store at -20°C for later use.

[0051] 4. RT-PCR amplification

[0052] 1) Primer design

[0053] The primers for PCR amplification were designed according to the gene coding sequence of TCONS_00016233 and Gapdh, and were synthesized by Guangzhou Ruqi Biotechnology Co.,...

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Abstract

A molecular marker, a kit using same, and an application thereof. The molecular marker is used for early diagnosis and prediction of sepsis complicated by acute kidney injury. The molecular marker in the blood of patients with sepsis complicated by acute kidney injury is highly expressed. Detecting expression levels of the molecular marker in the blood of a patient helps to determine whether the patient suffers from sepsis complicated by acute kidney injury or the probability of suffering from sepsis complicated by acute kidney injury.

Description

technical field [0001] The invention relates to the technical field of molecular diagnosis, in particular to a molecular marker, kit and application for early diagnosis and prediction of sepsis complicated with acute kidney injury. Background technique [0002] Sepsis is one of the common causes and complications of critically ill patients, and its case fatality rate is over 50%. At the same time, sepsis is also a common cause of acute kidney injury in critically ill patients. Once acute kidney injury occurs in patients with sepsis, the mortality rate can be as high as 70%. Therefore, early diagnosis and treatment are the key to reducing the morbidity and mortality of sepsis-related AKI. Currently, the diagnosis of acute kidney injury mainly relies on serum creatinine and urine output, which are affected by many factors and cannot reflect changes in renal function in a timely and accurate manner. They lack sufficient sensitivity and specificity for the diagnosis of AKI. It...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/68C12Q1/6883
Inventor 张东山瞿思源李惠玲
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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