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Kit for detecting versican V1 mRNA in urine and its application

A detection kit and detection reagent technology, applied in the field of medical detection, can solve the problems of poor long-term prognosis and severe clinical pathological damage, and achieve the effects of rapid and accurate detection of detection methods and convenient clinical application.

Active Publication Date: 2020-06-05
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although FSGS and minimal change disease (MCD) belong to the same podocyte disease, the clinicopathological damage is more severe than that of MCD, and the long-term prognosis is worse than that of MCD.

Method used

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  • Kit for detecting versican V1 mRNA in urine and its application
  • Kit for detecting versican V1 mRNA in urine and its application
  • Kit for detecting versican V1 mRNA in urine and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1: RT-PCR analysis of urinary sediment versican V1 mRNA

[0030] Collect 50ml of urine from the patient, centrifuge at 700g for 15min, and obtain urine sediment.

[0031] Total RNA was extracted using the Trizol method. The specific method is: add 1ml of Trizol to the sample, let stand at room temperature for 5 minutes, fully lyse, and completely separate the nucleoprotein complex. Add 1 / 5 of the volume of Trizol in chloroform, shake upside down for 15 seconds, let stand at room temperature for 5 minutes, and centrifuge (12000g, 15 minutes, 4°C). Transfer the upper aqueous phase to another clean EP tube, add an equal volume of isopropanol, let stand at room temperature for 5 minutes, and centrifuge (12000g, 10 minutes, 4°C). Remove the supernatant, add 1ml of 75% ethanol to wash the RNA pellet, mix with a shaker, and centrifuge (12000g, 5 minutes, 4°C); repeat the wash once; remove the supernatant, and centrifuge the empty tube (12000g, 1 minute, 4°C) . Remo...

Embodiment 2

[0041]PASM staining was used to evaluate renal tubular damage and interstitial fibrosis, and Masson staining area was used to semi-quantitatively analyze tubulointerstitial fibrosis. 50% was 3 points point. There was a significant correlation between the level of versican V1 mRNA in urinary sediment and the score of tubulointerstitial fibrosis in patients with FSGS, with a P value of figure 2 ).

Embodiment 3

[0043] The monthly decline rate of eGFR was calculated using a mixed linear effects model:

[0044] GFR_ij=beta0+beta1*time_ij+b_0i+b_1i*time_ij+epsilon_ij

[0045] eGFR decline rate = (beta1+b_1i) / (beta0+b_0i)

[0046] There was a significant correlation between the level of versican V1 mRNA in urinary sediment and the monthly decline rate of eGFR in patients with FSGS, with a P value of 0.001 ( image 3 ).

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Abstract

The invention belongs to the field of medical science detection, and especially relates to a kit for detecting versican V1mRNA in urine, and an application thereof. The urine of a patient is collected, and the versican V1mRNA level of urinary sediment is determined. A result shows that the versican V1mRNA level of the urinary sediment of an FSGS patient is significantly increased, and the versicanV1mRNA level of the urinary sediment of a minimal change disease (MCD) patient has no change; and the versican V1mRNA level of the urinary sediment of the FSGS patient is associated with the tubulointerstitial fibrosis score of the patient, and is significantly associated with the decline rate of eGFR during patient follow-up, so the expression level of the versican V1mRNA in urine can be used for the prognostic diagnosis of the FSGS patient. A reagent for detecting the versican V1mRNA expression level of urine can be used to prepare a detection reagent or the detection kit in order to realize the non-invasive diagnosis of the FSGS prognosis.

Description

technical field [0001] The invention belongs to the field of medical detection, in particular to a kit for detecting versican V1 mRNA in urine and its application in the prognosis diagnosis of focal segmental glomerulosclerosis. Background technique [0002] Focal Segmental Glomerular Sclerosis (FSGS) is a typical podocyte disease. Most of the patients have non-selective proteinuria, and hypertension and renal dysfunction are more common at the onset, often accompanied by renal tubular dysfunction. Pathologically, focal segmental lesions of glomeruli and acute and chronic lesions of tubulointerstitium can be seen in patients with FSGS. Although FSGS and minimal change disease (MCD) belong to the same podocyte disease, the clinicopathological damage is more severe than that of MCD, and the long-term prognosis is worse than that of MCD. The degree of chronic tubulointerstitial disease is closely related to the prognosis of FSGS. It is of great practical significance to find...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/118C12Q2600/158
Inventor 鲍浩刘志红韩润鸿秦卫松
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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