Application of miR-376b and detection kit

A technology of mir-376b and 1.mir-376b, which is applied in the field of acute kidney injury detection in patients with sepsis, can solve the problems of lack of specificity in diagnosis, and achieve the effects of high diagnostic sensitivity, non-degradation and mortality reduction.

Active Publication Date: 2020-03-13
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

However, these indicators still have significant limitations in the diagnosis of septic AKI
For example, in the absence of AKI, sepsis itself can also cause upregulation of NGAL, and the diagnosis lacks specificity

Method used

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  • Application of miR-376b and detection kit
  • Application of miR-376b and detection kit
  • Application of miR-376b and detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Gene chip analysis: AKI induced by intraperitoneal injection of lipopolysaccharide (LPS) in mice with sepsis, and microRNA expression profile changes in kidney tissue were analyzed by chip technology. The results showed that after 24 hours of LPS treatment, the expression of some microRNAs was up-regulated or down-regulated, including miR-376b. These results reveal the possible changing trend of microRNA in sepsis AKI and lay the foundation for the next experiments.

[0041] Animal experiment design:

[0042] C57BL / 6 mice, 6-8 weeks old, were purchased from Changsha Slake Jingda Experimental Animal Co., Ltd. We induced sepsis animal model by intraperitoneal injection (dose 10 mg / kg) of LPS (purchased from Sigma, USA). The specific grouping is shown in Table 1: (LPS was dissolved in normal saline, and the control group was simply injected with normal saline)

[0043] Table 1

[0044]

[0045] After the injection, the mice were put back into the breeding cage and p...

Embodiment 2

[0050] In addition to animal experiments, clinical specimens were collected at the same time, and relevant clinical tests and tests were carried out. A total of 40 urine samples from sepsis patients in our hospital (the Second Xiangya Hospital of Central South University) were collected; 20 cases of sepsis complicated with AKI, 20 cases of sepsis without AKI (control) (all sepsis cases The patients were previously healthy when they were collected, and some basic indicators and inflammatory indicators are shown in Table 2 below).

[0051] Table 2

[0052] sepsis non-AKI sepsis-AKI P value Gender: Male Female) 13 / 7 8 / 12 0.12 age 54.85±4.11 55.20±4.13 0.94 Serum creatinine, μmol / L 76.71±7.25 345.20±53.63 <0.01*

[0053] Total RNA was extracted from the patient's urine sample, cDNA was synthesized using taqman probe (human-derived miR-376b probe, primer catalog number: 001102, taqman microRNA Reverse Transcirculation Kit, all purchased from ...

Embodiment 3

[0055] We used receiver operating characteristic curve (ROC) to evaluate the validity of urinary miR-376b as a diagnostic indicator of sepsis AKI. The result is as Figure 6 , AUC=0.81, which has good diagnostic value. According to the ROC curve, we believe that the diagnostic effect is the best when the Youden index (Youden index=sensitivity+specificity-1) is maximum, and the diagnostic cut-off value determined according to this principle is urinary miR-376b<0.69 times. The final diagnostic cut-off values ​​and their diagnostic sensitivity and specificity are shown in Table 3.

[0056] table 3

[0057]

[0058] Urinary [TIMP2]*[IGFBP7] is a sensitive indicator approved by the US FDA for the early diagnosis of AKI, and its diagnostic value in AKI has been widely recognized. The present invention uses the same sample to measure urine [TIMP2]*[IGFBP7], and the ROC curve of urine [TIMP2]*[IGFBP7] to diagnose sepsis AKI is shown in Figure 7 , indicating that compared with ...

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Abstract

The invention discloses the application of miR-376b and a detection kit. The invention specifically relates to the application of a reagent for detecting miR-376b in the preparation of preparations used for predicting acute kidney injuries in human beings or mice suffering from sepsis. In the body of a patient with sepsis and acute kidney injuries, the expression of the urine miR-376b is reduced,and a blood creatinine value or a urea nitrogen value has negative correlation with the expression of the urine miR-376b, which prompt that down-regulation of miR-376b expression may be related to acute kidney injuries of patients with sepsis. Results show that miR-376b can be used as a biomarker for AKI diagnosis of sepsis, and has the advantages of early diagnosis and high sensitivity and specificity.

Description

technical field [0001] The invention belongs to the technical field of detecting acute kidney injury in patients with sepsis, and specifically relates to the application of a reagent for detecting miR-376b in the preparation of preparations for predicting acute kidney injury in patients with sepsis, and a detection kit. Background technique [0002] Acute kidney injury (acute kidney injury, AKI), also known as acute renal failure, is a rapid decline or even loss of kidney function caused by a variety of reasons, resulting in the body not being able to excrete various toxins and metabolites in the body in time, which in turn causes various Harmful substances or normal metabolites in the body accumulate in the body, causing disturbance of the internal environment and abnormal metabolism of the body, and symptoms of various systems of the body appear. Clinically, AKI is mainly caused by sepsis, ischemia-reperfusion (IR) injury and various endogenous and exogenous nephrotoxic su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 董政刘志文唐程远高婧丽
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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