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Markers for diagnosing and predicating focal segmental glomerulosclerosis

A glomerulosclerosis and focal segmental technology, applied in the field of detection, can solve the problems of lack of blood diagnostic markers, etc., and achieve good market prospects, low detection cost, and high sensitivity

Inactive Publication Date: 2016-07-20
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main diagnosis of FSGS is based on renal biopsy, and there is a lack of specific blood diagnostic markers

Method used

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  • Markers for diagnosing and predicating focal segmental glomerulosclerosis
  • Markers for diagnosing and predicating focal segmental glomerulosclerosis
  • Markers for diagnosing and predicating focal segmental glomerulosclerosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1, Taqman real-time quantitative PCR detection plasma miRNAs expression profile

[0019] Take the plasma of 20 cases of clinically diagnosed FSGS patients, and take the serum of 20 cases of healthy people as a control, and then extract plasma RNA, the extraction method refers to mirVana TM The PARIS kit is operated, and the specific steps are as follows:

[0020] ①Add an equal volume of 2×DenaturingSolution to 400 μL plasma, vortex for 30 seconds, mix thoroughly, add 10 μL cel-mir-39 (10 nM), and immediately place on ice for 5 minutes;

[0021] ② Add 800 μL of phenol chloroform, vortex for 30-60 seconds;

[0022] ③Centrifuge at 12000×g, 4°C for 15 minutes, separate the supernatant into a clean centrifuge tube;

[0023] ④ Add 1.25 times the volume of the supernatant and 100% ethanol at room temperature to the separated supernatant, and mix well;

[0024] ⑤Pour the mixed liquid into the spin column. If the liquid volume is greater than 700 μL, load it on the...

Embodiment 2

[0033] Example 2, Candidate marker verification for FSGS diagnosis

[0034] In order to verify whether the four candidate markers screened in Example 1 can be used for early diagnosis and prognosis of FSGS, this example expands the sample size. Get 60 routine plasma samples (30 routine FSGS patient plasma and 30 routine healthy person plasma), carry out real-time quantitative PCR according to the method for embodiment 1, the result is as follows figure 2 shown. The results showed that the expressions of miR-17, miR-451, miR-106a and miR-19b in the plasma of FSGS patients were significantly reduced in 60 samples, and it was preliminarily determined that miR-17, miR-451, miR-106a and miR-19b could be used as Markers for early diagnosis and prognosis of FSGS.

[0035] Then the ROC curve was drawn according to the detection results, and the diagnostic sensitivity and specificity of miR-17, miR-451, miR-106a and miR-19b were judged by the area under the curve (AUC) (the closer t...

Embodiment 3

[0036] Example 3, clinical application of plasma miRNAs markers in the diagnosis of FSGS

[0037] In order to evaluate the effect of miR-17, miR-451, miR-106a and miR-19b as markers in the clinical diagnosis of FSGS, 30 plasma specimens of FSGS confirmed by renal tissue biopsy were collected, and 30 healthy human plasma samples were collected as controls , perform quantitative PCR according to the method of Example 1, respectively detect the expression of miR-17, miR-451, miR-106a and miR-19b, and use the 4 miRNAs markers alone or in combination to predict the incidence probability of FSGS. The results show that when miR-17 is detected alone, 23 cases of FSGS can be diagnosed in 30 cases of confirmed FSGS patients, and its sensitivity is 76.7%. Among 30 cases of normal plasma, 24 cases can be diagnosed as normal, and its specificity 80.0%; when miR-451 is detected alone, 25 cases of FSGS can be diagnosed in 30 cases of confirmed FSGS patients, and its sensitivity is 83.3%. In ...

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Abstract

The invention discloses application of reagent of markers for detecting focal segmental glomerulosclerosis (FSGS) to preparation of a kit for diagnosing and predicating focal segmental glomerulosclerosis, the focal segmental glomerulosclerosis markers are one or more of miR-17, miR-451, miR-106a and miR-19B, By means of the markers for diagnosing and predicating focal segmental glomerulosclerosis, new blood markers are provided for clinical diagnosis of focal segmental glomerulosclerosis, peripheral blood testing can be directly conducted, and the markers have the advantages of being good in specificity and high in sensitivity, have good clinical diagnosis value and lay a foundation for implementation of hematology diagnosis of FSGS.

Description

technical field [0001] The invention belongs to the field of detection, in particular to the application of reagents for detecting markers of focal segmental glomerulosclerosis in the preparation of reagent kits for diagnosing and predicting focal segmental glomerulosclerosis. Focal segmental glomerulosclerosis The marker of glomerulosclerosis is at least one of miR-17, miR-451, miR-106a and miR-19b. Background technique [0002] Focal segmental glomerulosclerosis (FSGS) is a common primary glomerular disease, the incidence of which has gradually increased in recent years, and about 30% to 40% of adult patients with primary nephrotic syndrome for FSGS. The clinical manifestations are non-specific, and most of them start with insidious nephrotic syndrome. If not diagnosed and treated in time, they will develop into end-stage renal disease. Currently, the main diagnostic basis for FSGS is renal biopsy, which lacks specific blood diagnostic markers. Therefore, there is an ur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/178
Inventor 肖斌邹全明汪莉娜赵洪雯龚莉余婷
Owner ARMY MEDICAL UNIV
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