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Kit for predicting prostate screening and lymphatic metastasis

A technology for prostate cancer and lymphatic metastasis, applied in the field of biomedicine, can solve the problems of inability to distinguish other tumors, the methylation site is not specific enough, and insufficient to distinguish prostatic hyperplasia and prostatitis, and achieves high clinical positive predictive value and negative Predictive value and detection result are accurate and reliable

Inactive Publication Date: 2018-05-04
上海纽思格生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the methylation sites reported by such scientific research are not specific enough to distinguish other tumors such as bladder cancer, nor are they sufficient to distinguish benign prostatic hyperplasia and prostatitis, so there are no formed products so far

Method used

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  • Kit for predicting prostate screening and lymphatic metastasis
  • Kit for predicting prostate screening and lymphatic metastasis
  • Kit for predicting prostate screening and lymphatic metastasis

Examples

Experimental program
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Effect test

Embodiment example 1

[0037] Implementation Case 1: Urine from 50 inpatients (not less than 10ml) from the Urology Department of Shanghai Tenth People’s Hospital, including 25 bladder cancer patients, 11 prostate patients, and the rest are non-tumor patients of the urinary system such as benign prostatic hyperplasia, prostatic hyperplasia Inflammation, urethritis and other patients. Through the technical scheme of the present invention, the following primer-probe combinations are used for detection: KDM5D (FAM fluorescence), EFEMP1 (VIC fluorescence), PITX2 (ROX fluorescence), RASSF1 (CY5 fluorescence), GSTP1 (NED fluorescence), all using high hybridization For MGB probes with high resolution, run the program on ABI7500: 50°C for 2min, 95°C for 5min, 95°C for 10sec, 60°C for 40sec, cycle 40-45 times, 5-color fluorescence detection. In addition, the detection of monochromatic fluorescence (both labeled with FAM) can achieve the same effect, but the nucleic acid template needs to be detected in sever...

Embodiment example 2

[0059] Example 2: 38 urine samples (not less than 10ml) from the Urology Department of Shanghai Pudong Hospital, including 9 bladder cancer patients, 20 prostate patients, and 9 non-tumor patients in the urinary system. Through the technical scheme of the present invention, use the following primer probe combinations to detect, CRMP4 (FAM), WFDC2 (VIC), TACSTD2 (NED), CYBA (CY5), all adopt the MGB probe of high hybridization resolution, run on ABI7500 Program: 50°C for 2min, 95°C for 5min, 95°C for 10sec, 60°C for 40sec, cycle 40-45 times, 4-color fluorescence detection.

[0060] Test results such as Figure 2-1 to Figure 2-4 : 18 out of 20 prostate patients were detected positive, while 9 bladder cancer patients and 9 non-tumor patients were all negative. The positive predictive value is 100%, and the negative predictive value is 90%. In addition, lymph node metastasis was predicted according to the positive detection of CRMP4: 8 cases had lymph node metastasis in 18 cases,...

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Abstract

The invention relates to the molecular diagnosis field and relates to a molecular diagnosis method and reagent for non-invasive detection of prostate cancer and lymphatic metastasis of prostate cancerby virtue of urine. The method comprises the following steps: extracting and purifying urine nucleic acid; (2) carrying out DNA hydrosulphite conversion reaction and subsequent purification; (3) detecting a plurality of gene promoter regions by virtue of methylation-specific real-time fluorescence PCR; and (4) analyzing multiple gene results, wherein the steps (1) and (2) can be combined into onestep, and multiple genes contain the promoter regions of the following genes: CRMP4, PITX2, RASSF1A, SOSTDC, CYBA, EFEMP1, KNM5D, GSTP1, WFDC2, TACSTD2 and the like. By combining the detection results of the genes, the diagnosis of prostate cancer and the lymphatic metastasis of prostate cancer can be realized.

Description

technical field [0001] The present invention belongs to the field of biomedicine, especially the field of molecular diagnosis related to non-invasive tumor screening. Development related, including detection, prognosis, lymph node metastasis, etc. Background technique [0002] Prostate cancer is one of the common male reproductive system malignancies. Worldwide, the incidence of prostate cancer ranks second among all malignant tumors in men. Prostate cancer (PCa) is growing faster than other malignancies in China due to lifestyle changes and a declining proportion of the aging population. At present, the clinical diagnosis methods of prostate cancer mainly include serum prostate-specific antigen (PSA) detection, digital rectal examination, rectal ultrasonography, biopsy pathological examination, etc. Prostate-specific antigen (PSA) is widely used as a molecular marker for clinical diagnosis, but in some cases or physiological conditions, PSA will enter the blood, resultin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6806C12Q1/686C12N15/10
CPCC12Q1/6886C12N15/1013C12Q1/6806C12Q1/686C12Q2600/154C12Q2563/107C12Q2561/101
Inventor 周科隆宋军
Owner 上海纽思格生物科技有限公司
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