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miRNA (microribonucleic acid) biomarkers and detection kit for ovarian cancer diagnosis

A biomarker and ovarian cancer technology, applied in the detection/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of shallow understanding of miRNA and urgent need for further investigation of ovarian cancer

Inactive Publication Date: 2015-12-23
崔长友
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current understanding of miRNA self-regulation mechanism is still relatively shallow, and its research in ovarian cancer is urgently needed.

Method used

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  • miRNA (microribonucleic acid) biomarkers and detection kit for ovarian cancer diagnosis
  • miRNA (microribonucleic acid) biomarkers and detection kit for ovarian cancer diagnosis
  • miRNA (microribonucleic acid) biomarkers and detection kit for ovarian cancer diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Screening of differential miRNAs in ovarian cancer tissues

[0022] 1 Objects and methods

[0023] 1.1 Specimen source

[0024] After obtaining the patient’s informed consent, 8 pairs of postoperative specimens from patients with ovarian cancer confirmed by pathology in Jiangsu Provincial People’s Hospital from April 2013 to June 2014 were collected, including cancer tissue specimens and paired paracancerous tissues that were more than 3 cm away from the cancer tissue. All patients had not received chemotherapy and radiotherapy before operation.

[0025] 1.2 Main instruments and equipment

[0026] Desktop centrifuge: Eppendorf Minispin (USA) Eppendorfcentrifuge5810R (USA)

[0027] Nucleic acid concentrator: Eppendorfconcentrator5301 (USA)

[0028] Ultraviolet spectrophotometer: DU640, balance (Beckman company product)

[0029] Ultraviolet crosslinking instrument: GSGENELINKERUVChamber (product of BIO-RAD company)

[0030] Water bath (product of Memert c...

Embodiment 2

[0100] Example 2: Serological analysis of differential miRNAs in ovarian cancer tissues

[0101] 1 Objects and methods

[0102] 1.1 Specimen source

[0103] After obtaining the informed consent of the patients, 6ml of fasting venous blood in the morning from May 2013 to March 2014 was collected from 165 patients with pathologically diagnosed ovarian cancer and 120 healthy people in Jiangsu Provincial People's Hospital as a control. All ovarian 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.

[0104] 1.2 Serum sample collection and processing

[0105] 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 serum and divide it into RNase-freeEP tubes ...

Embodiment 3

[0154] Embodiment 3: receiver operating characteristic curve (ROC) analysis

[0155] A ROC curve was constructed to compare the diagnostic ability of five serum miRNAs to distinguish ovarian cancer patients from healthy controls. The areas under the ROC curve (AUC) of the five miRNAs were: has-miR-193b-3p, 0.840 (95% confidence interval: 0.760-0.920); has-miR-155-5p, 0.819 (95% confidence interval: 0.736-0.902 ); has-miR-145-5p, 0.814 (95% confidence interval: 0.734-0.894); has-miR-132-3p, 0.843 (95% confidence interval: 0.765-0.920); has-miR-143-3p, 0.738 (95% confidence interval: 0.644-0.832). Under the optimal cutoff value, the sensitivity and specificity of miRNA are as follows: has-miR-193b-3p, respectively 83.3% and 82.7%; has-miR-155-5p, respectively, 64.8% and 94.2%; has- miR-145-5p, respectively 90.7% and 61.5%; has-miR-132-3p, respectively, 70.4% and 82.5%; has-miR-143-3p, respectively, 67.3% and 74.1%. The combined AUC of these five miRNAs can reach 0.973, and th...

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Abstract

The invention discloses miRNA (microribonucleic acid) biomarkers and a detection kit for ovarian cancer diagnosis. The microRNA biomarkers are composed of the following microRNAs: has-miR-193b-3p, has-miR-155-5p, has-miR-145-5p, has-miR-132-3p and has-miR-143-3p. The serological expression analysis on the five miRNAs has-miR-193b-3p, has-miR-155-5p, has-miR-145-5p, has-miR-132-3p and has-miR-143-3p of ovarian cancer with obvious para-carcinoma tissue expression differences (the differential expression level is greater than 2 folds, and the CT value in RT-PCR (reverse transcription-polymerase chain reaction) is less than 30) indicates that the five miRNAs have stable expression in serum, the serum miRNA expression has favorable consistency with the tissue, the expressions of the has-miR-193b-3p, has-miR-155-5p and has-miR-145-5p are enhanced, and the expressions of the has-miR-132-3p and has-miR-143-3p are lowered. The five miRNAs can be used as biomarkers for ovarian cancer diagnosis, and the sensitivity and specificity of combined diagnosis are obviously higher than the sensitivity and specificity 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 ovarian cancer. Background technique [0002] Ovarian cancer is one of the three common malignant tumors of the female reproductive system, and its incidence rate ranks third. However, due to the lack of effective early diagnosis methods and the characteristics of ovarian cancer resistance to chemotherapy and easy metastasis and recurrence, the mortality rate of ovarian cancer ranks first among gynecological malignancies, and the 5-year survival rate is less than 30%, which seriously threatens the life and health of women. Although studies have discovered various molecular signaling pathways related to malignant transformation and drug resistance of epithelial ovarian cancer (EOC), how to make early diagnosis and predict response to chemotherapy is still a major challenge in the clinical treatment of EOC. [0003] MicroRN...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 崔长友
Owner 崔长友
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