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Diagnostic agent for diagnosis or auxiliary diagnosis of renal insufficiency or renal injury, kit and application

A technology of auxiliary diagnosis and diagnostic agent, applied in the direction of disease diagnosis, biological testing, biochemical equipment and methods, etc., can solve the problems of lack of drug treatment and unclear pathogenic mechanism

Active Publication Date: 2020-03-24
SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although renal IRI is an important cause of morbidity and death in AKI patients, its specific pathogenic mechanism is still unclear and there is a lack of effective drug treatment

Method used

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  • Diagnostic agent for diagnosis or auxiliary diagnosis of renal insufficiency or renal injury, kit and application
  • Diagnostic agent for diagnosis or auxiliary diagnosis of renal insufficiency or renal injury, kit and application
  • Diagnostic agent for diagnosis or auxiliary diagnosis of renal insufficiency or renal injury, kit and application

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Experimental program
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Effect test

Embodiment 1

[0040] Proteomics label-free quantitative technology (Label-free) is an important qualitative and quantitative method of mass spectrometry in recent years. After the extracted protein is enzymatically hydrolyzed, the peptides are analyzed by mass spectrometry using liquid chromatography-mass spectrometry. By comparing the corresponding peptides in different samples The signal intensity of the peptide can be used to perform relative quantification of the protein corresponding to the peptide.

[0041] Experimental principle: the abundance (quantity) of each peptide is proportional to its peak area or signal intensity in MS1; while the retention time (RT) of the same peptide in different samples on LC is consistent, Therefore, quantitative analysis of the relative expression levels of proteins in different samples can be achieved by comparing the MS spectra and then integrating them into the corresponding proteins.

[0042] In this embodiment, the HK2 human renal tubular epitheli...

Embodiment 2

[0044] HE and PAS staining to detect renal injury

[0045] IRI model: After C57 mice were anesthetized, the left renal pedicle was separated, and the left renal artery and vein were clamped with a non-invasive microvascular clip to block the blood flow of the left kidney, and placed in a 32 degree Celsius incubator. blood at different times (0, 15, 30, 45, 60, 120, 240, 360 min), and then loosen the left renal vascular clamp to restore the blood perfusion of the left kidney. The mice were killed 24 hours after the blood perfusion of the kidney was restored, and the kidney tissue was collected .

[0046] Hematoxylin-eosin staining (HE) staining and Periodic Acid-Schiff (PAS) staining, HE and PAS are common paraffin pathological staining techniques, often used in renal pathology Assessment of sexual injury.

[0047] The principle of HE staining method: the hematoxylin staining solution is alkaline, which mainly makes the chromatin in the nucleus and the nucleic acid in the cyt...

Embodiment 3

[0051] Expression of SRP14 and RPS7 detected by immunohistochemistry

[0052] IRI model: After C57 mice were anesthetized, the left renal pedicle was separated, and the left renal artery and vein were clamped with a non-invasive microvascular clip to block the blood flow of the left kidney, and placed in a 32 degree Celsius incubator. blood at different times (0, 15, 30, 45, 60, 120, 240, 360 min), and then loosen the left renal vascular clamp to restore the blood perfusion of the left kidney. The mice were killed 24 hours after the blood perfusion of the kidney was restored, and the kidney tissue was collected and serum. The expression of SRP14 and RPS7 in the paraffin-embedded kidney tissue sections of mice was detected by immunohistochemistry. Immunohistochemical method: According to the principle of antigen-antibody reaction and chemical color development, the antigen in the mouse kidney tissue section is first combined with the primary antibody of SRP14 or RPS7, and then...

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Abstract

The invention discloses a diagnostic agent for diagnosis or auxiliary diagnosis of renal insufficiency or renal injury, a kit and application, and relates to the technical field of renal insufficiencyor renal injury diagnosis. The diagnostic agent disclosed by the invention comprises a detection reagent for detecting the expression level of RPS7 gene and / or the SRP14 gene. Researches find that the RPS7 gene and the SRP14 gene have the characteristic of high expression in a renal insufficiency or renal injury group, can be used as markers for renal insufficiency or renal injury diagnosis or auxiliary diagnosis, and provide a new thought and method for renal insufficiency or renal injury diagnosis or auxiliary diagnosis.

Description

technical field [0001] The present invention relates to the technical field of diagnosis of renal insufficiency or renal injury, in particular to a diagnostic agent, kit and application for diagnosis or auxiliary diagnosis of renal insufficiency or renal injury. Background technique [0002] Renal insufficiency or acute kidney injury (AKI) can easily progress to chronic kidney disease (CKD) and end stage renal disease (ESRD), with a mortality rate as high as 50%. In recent years, the morbidity and fatality rate of this disease have been increasing continuously, causing great harm to families and society. Renal ischemia reperfusion injury (renal ischemia reperfusion injury, renal IRI) is an important cause of AKI, and also one of the main factors affecting the early functional recovery and long-term survival of the transplanted kidney after renal transplantation. The kidney is one of the organs sensitive to ischemia-reperfusion injury due to its special structure and functio...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/6893G01N2333/47G01N2800/347
Inventor 李怡
Owner SICHUAN PROVINCIAL PEOPLES HOSPITAL
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