Application of exosome gene, prostate cancer detector, and detection kit and detection device thereof

A detection kit and prostate cancer technology, applied in the field of genetic engineering, can solve the problems of low positive detection rate, large error, elevated PSA, etc.

Active Publication Date: 2021-08-13
深圳市拾方杰科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Serum PSA is currently used as a clinical detection marker for prostate cancer, and its specificity is not high. About 30% of PCa patients may not increase serum PSA, but fluctuate within the normal range (normal range<4.0ng / ml), about 40% of normal healthy individuals detect elevated PSA, with large errors and a low positive detection rate. PSA has been questioned due to excessive puncture and overdiagnosis caused by the "PSA-based PCa screening strategy"
The early diagnosis method of prostate cancer, which mainly relies on imaging and pathological biopsy, has a high detection rate, but because prostate cancer has the characteristics of hidden symptoms, most patients are already in the middle and late stages when they are diagnosed, but the cure rate of early prostate cancer can reach 80%. , while the cure rate of advanced prostate cancer is only 10% to 20%, and cell and histopathological detection requires the use of puncture or surgery to obtain tumor tissue samples from patients, which is not only inconvenient, but also causes harm to patients. bodily injury

Method used

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  • Application of exosome gene, prostate cancer detector, and detection kit and detection device thereof
  • Application of exosome gene, prostate cancer detector, and detection kit and detection device thereof
  • Application of exosome gene, prostate cancer detector, and detection kit and detection device thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Extraction and Identification of Urine Exosomes

[0072] After massaging the prostate, collect 20 mL of morning urine, store at 4°C, centrifuge at 2000g for 10 minutes to remove residual cells and collect the supernatant, and then centrifuge at 10000g for 10 minutes to remove cell debris.

[0073] Take 10mL of the supernatant to a new centrifuge tube, add exosome extraction reagent 1 (Shenzhen Shifangjie Technology Co., Ltd.) with a volume ratio of 2:1; invert and mix well, let stand at 4°C for 5min, and then centrifuge at 10,000g 2min; take the supernatant and transfer it to a new centrifuge tube, add exosome extraction reagent 2 (Shenzhen Shifangjie Technology Co., Ltd.) with a volume ratio of 1:1; after inverting and mixing, stand at 4°C for 1 hour After centrifugation at 12000g for 30 min, the supernatant was discarded; the precipitate was collected, and 0.5 mL of PBS buffer was added to resuspend the precipitate, and the exosome suspension was prepared by blowing a...

Embodiment 2

[0076] Exosome gene detection

[0077] 2.1 Extraction of urine exosome RNA:

[0078] ①Take 0.5mL of exosome resuspension and put it into a 1.5ml RNA-free centrifuge tube, add 1mL Trizol lysate, vortex and mix, and let stand at room temperature for 5mins;

[0079] ② Add 200 μL chloroform, shake vigorously for 15 seconds, and let stand at room temperature for 2 minutes;

[0080] ③ After centrifugation at 12000×g for 15 minutes at 4°C, divide into three layers, transfer the upper aqueous phase to a new 1.5mL RNA-free EP tube, be careful not to touch the white membrane in the middle layer;

[0081] ④ Add pre-cooled 0.6mL isopropanol to the tube, mix well, and let stand at room temperature for 10mins;

[0082] ⑤Centrifuge at 12000×g for 10mins at 4°C, discard the supernatant, and sink the RNA to the bottom of the tube;

[0083] ⑥ Add 1 mL of pre-cooled 75% ethanol diluted with sterilized DEPC water, suspend the precipitate, shake gently, and wash the RNA;

[0084] ⑦ Centrifuge ...

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Abstract

The invention relates to application of an exosome gene, a prostate cancer detector, and a detection kit and a detection device of the prostate cancer detector. The application of the exosome gene as a biomarker in preparation of a prostate cancer detection reagent, a prostate cancer detection kit or a prostate cancer detection device is characterized in that the exosome gene contains at least one of a PCA3 gene, an SPDEF gene and a TMPRSS2: ERG fusion gene. The biomarker is used for preparing a prostate cancer detection reagent, a prostate cancer detection kit or a prostate cancer detection device, and the positive detection rate of prostate cancer is high.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to an application of exosome genes, a prostate cancer detection substance, a detection kit and a detection device thereof. Background technique [0002] Prostate cancer (PCa) is the most common malignant tumor of the urinary system worldwide and the fifth leading cause of cancer death in men. It is also the malignant tumor with the fastest increasing incidence and mortality in China in recent years. Early diagnosis of cancer is a diagnosis and treatment method specifically for early cancer patients. Its purpose is early detection and early treatment, so as to reduce the pain, mental and economic burden of patients, and strive for early recovery of cancer patients through early diagnosis and treatment of cancer. The clinical diagnosis of prostate cancer mainly relies on digital rectal examination, serum PSA, transrectal ultrasound of the prostate and pelvic MRI. The diagnosis of pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11G16B20/00G16H50/20
CPCC12Q1/6886G16B20/00G16H50/20C12Q2600/158C12Q2600/166
Inventor 刘好罗敏虹
Owner 深圳市拾方杰科技有限公司
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