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miRNA biomarker and detection kit used for renal cancer diagnosis

A technology of biomarkers and kits, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve problems such as microRNA biomarkers that have not been found in kidney cancer

Inactive Publication Date: 2016-07-20
崔学俊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] To date, no reliable microRNA biomarkers have been identified for the diagnosis of kidney cancer

Method used

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  • miRNA biomarker and detection kit used for renal cancer diagnosis
  • miRNA biomarker and detection kit used for renal cancer diagnosis
  • miRNA biomarker and detection kit used for renal cancer diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Screening of differential miRNAs in renal cancer tissues

[0016] 1 Objects and methods

[0017] 1.1 Specimen source

[0018] After obtaining the patient’s informed consent, 8 pairs of specimens were collected from Jiangsu Provincial Hospital of Integrated Traditional Chinese and Western Medicine from April 2013 to April 2014, which were confirmed by pathology as kidney cancer patients after surgery, including cancer tissue specimens and paired specimens with a distance of more than 3 cm from the cancer tissue. Paracancerous tissue, all patients had not received chemotherapy and radiotherapy before operation.

[0019] 1.2 Main instruments and equipment

[0020] Desktop centrifuge: Eppendorf Minispin (USA) Eppendorfcentrifuge5810R (USA)

[0021] Nucleic acid concentrator: Eppendorfconcentrator5301 (USA)

[0022] Ultraviolet spectrophotometer: DU640, balance (Beckman company product)

[0023] Ultraviolet crosslinking instrument: GSGENELINKERUVChamber (prod...

Embodiment 2

[0094] Example 2: Serological Analysis of Differential miRNAs in Renal Cancer Tissues

[0095] 1 Objects and methods

[0096] 1.1 Specimen source

[0097] After obtaining the informed consent of the patients, 165 patients with pathologically diagnosed kidney cancer and 120 healthy people with early morning fasting venous blood (6ml) were collected from Jiangsu Provincial Hospital of Integrated Traditional Chinese and Western Medicine from May 2013 to August 2014 as a control. All kidney cancer patients were diagnosed for the first time, and had not undergone surgery, radiotherapy or chemotherapy before blood collection. The 120 healthy control groups were age-matched healthy people without malignant tumors or other diseases.

[0098] 1.2 Serum sample collection and processing

[0099] Draw 6ml of fasting venous blood in the morning and put it in a tube without anticoagulant, let it stand for 30 minutes, centrifuge at 1300g (or 4000rpm) at 4°C for 15 minutes, take the upper ...

Embodiment 3

[0149] Embodiment 3: receiver operating characteristic curve (ROC) analysis

[0150] A ROC curve was constructed to compare the diagnostic ability of five serum miRNAs in distinguishing RCC patients from healthy controls. The areas under the ROC curve (AUC) of the five miRNAs were: hsa-miR-10b-5p, 0.796 (95% confidence interval: 0.760-0.920); hsa-miR-384, 0.789 (95% confidence interval: 0.736-0.902); hsa-miR-206, 0.774 (95% confidence interval: 0.734-0.894); hsa-miR-424-5p, 0.793 (95% confidence interval: 0.765-0.920); hsa-miR-224-5p, 0.732 (95% Confidence interval: 0.644-0.832). Under the optimal cutoff value, the sensitivity and specificity of miRNAs are as follows: hsa-miR-10b-5p, 83.3% and 82.7%, respectively; hsa-miR-384, 64.8% and 94.2%, respectively; hsa-miR- 206, respectively, 90.7% and 61.5%; hsa-miR-424-5p, respectively, 70.4% and 82.5%; hsa-miR-224-5p, respectively, 67.3% and 74.1%. The combined AUC of these five miRNAs can reach 0.992, and the sensitivity and sp...

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Abstract

The invention discloses a miRNA biomarker and a detection kit which are used for renal cancer diagnosis. The microRNA biomarker is formed by the following microRNAs: hsa miR 10b 5p, hsa miR 384, hsa miR 206, hsa miR 424 5p and hsa miR 224 5p. According to the invention, serology expression analysis is carried out on five miRNAs obviously in differential expression of renal cancer and para-carcinoma tissues (differential expression quantity is more than 2 folds, and CT value in RT PCR is smaller than 30) hsa miR 10b 5p, hsa miR 384, hsa miR 424 5p, hsa miR 206 and hsa miR 224 5p, results show that the five miRNAs are stably expressed in serum, consistency between expression of serum miRNA and tissues is good, hsa miR 10b 5p, has miR 384 and has miR 206 are in down-regulated expression, and has miR 424 5p and has miR 224 5p are in up-regulated expression. The five miRNAs can serve as a biomarker used for the renal cancer diagnosis, and the sensitivity and specificity of combined diagnosis are obviously higher than those of single miRNA diagnosis.

Description

technical field [0001] The invention relates to biological detection, in particular to a microRNA biomarker and a detection kit for the diagnosis of human kidney cancer. Background technique [0002] Renal cell carcinoma (referred to as renal cell carcinoma) accounts for about 2%-3% of malignant tumors in adults, and is the second most common malignant tumor of the urinary system in my country, second only to bladder cancer. Surgical resection is the most effective treatment for localized RCC, but many patients eventually relapse. The vast majority of renal cell carcinomas have no clinical symptoms in the early stage, and some patients have locally advanced or distant metastases when they are first diagnosed, and cannot be radically removed by surgery, and the prognosis of these patients is poor. Therefore, early diagnosis of kidney cancer becomes particularly necessary. [0003] MicroRNA (miRNA) is a newly discovered non-coding small molecule RNA with a length of about 18...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 崔学俊
Owner 崔学俊
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