Analytical method for identifying prostate cancer based on urine exosomes

A prostate cancer and analysis method technology, which is applied in the analysis field of prostate cancer identification based on urine exosomes, can solve the problems of low accuracy rate, high re-examination rate, incompatibility of PCR instruments, etc., and achieve the effect of high accuracy rate

Inactive Publication Date: 2020-01-03
赵凯
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method improves the accuracy of the predictive diagnosis of prostate cancer and does not require biopsy to reduce the psychological burden of patients, the accuracy of quality control and results is low, and the retest rate is high, which is not compatible with conventional clinical PCR instruments. Need to purchase additional equipment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] An analysis method for identifying prostate cancer based on urine exosomes, comprising the steps of:

[0034] S1. Collect 10 mL of urine from the patient at the initial stage, centrifuge at 4°C and 2500 rpm for 5 minutes, discard the supernatant, add 1.0 mL of PBS solution to the bottom sediment, and repeatedly blow and mix to obtain a mixed exosome solution;

[0035] S2. Add 1.3mL Trizol solution to the exosome solution in S1, let stand for 3min, add 0.2mL chloroform, mix upside down, centrifuge at 2°C and 12000rpm for 13min, take the upper clear solution, and pass the upper clear solution through several centrifuges, Obtain sample RNA solution;

[0036] S3. Take 5 μL each of the positive quality control product, the negative quality control product, and the sample RNA solution in S2, respectively, and mix them with RT-PCR amplification reagents (the specific RT-PCR amplification reagents include PSA, PCA3, ERG, and SPDEF). Primers and probes) were amplified to obtain...

Embodiment 2

[0057] An analysis method for identifying prostate cancer based on urine exosomes, comprising the steps of:

[0058] S1. Collect 12 mL of urine from the patient at the initial stage, centrifuge at 2°C and 2200 rpm for 7 minutes, discard the supernatant, add 1.2 mL of LPBS solution to the bottom sediment, and repeatedly blow and mix to obtain a mixed exosome solution;

[0059] S2. Add 1.0mL Trizol solution to the exosome solution in S1, let it stand for 4min, add 0.4mL chloroform, mix it upside down, centrifuge at 4°C and 11000rpm for 12min, take the upper clear solution, and pass the upper clear solution through several centrifuges, Obtain sample RNA solution;

[0060] S3. Take 6 μL each of the positive quality control product, the negative quality control product, and the sample RNA solution in S2, and mix them with RT-PCR amplification reagents (the specific RT-PCR amplification reagents include PSA, PCA3, ERG, and SPDEF). Primers and probes) were amplified to obtain positi...

Embodiment 3

[0081] An analysis method for identifying prostate cancer based on urine exosomes, comprising the steps of:

[0082] S1. Collect 14mL of urine from the patient at the initial stage, centrifuge at 4°C and 2000rpm for 10min, discard the supernatant, add 1.4mL of PBS solution to the bottom sediment, and repeatedly blow and mix to obtain a mixed exosome solution;

[0083] S2. Add 1.5mL Trizol solution to the exosome solution in S1, let it stand for 5min, add 0.6mL chloroform, mix it upside down, centrifuge at 4°C and 11000rpm for 15min, take the upper clear solution, and pass the upper clear solution through multiple centrifuges, Obtain sample RNA solution;

[0084] S3. Take 7 μL each of the positive quality control product, the negative quality control product, and the sample RNA solution in S2, respectively, and mix them with RT-PCR amplification reagents (the specific RT-PCR amplification reagents include PSA, PCA3, ERG, and SPDEF). Primers and probes) were amplified to obtain...

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PUM

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Abstract

The invention discloses an analytical method for identifying prostate cancer based on urine exosomes in the field of prostate cancer. According to the analytical method for identifying prostate cancerbased on the urine exosomes,the exosomes are directly extracted from human urine, whether the prostate cancer is diagnosed or not is identified by measuring the content of PCA3, ERG group, and SPDEFin the exosomes,the accuracy of the identification result is higher than that of a urine detection kit adopting PCA3 / PSA, and the detection can be completed by using only ordinary PCR instruments without the need to purchase additional equipment.

Description

technical field [0001] The invention relates to the field of prostate cancer, in particular to an analysis method for identifying prostate cancer based on urine exosomes. Background technique [0002] Prostate cancer is the most common malignant tumor of the urinary system. With the aging population in my country, the incidence of prostate cancer is increasing year by year, which seriously threatens the life and health of elderly men. At present, the medical identification of prostate cancer is mainly through biopsy to detect the content of the patient's prostate specific antigen (PSA). PSA has good prostate tissue specificity, but lacks sufficient prostate cancer specificity. In patients with benign prostatic hyperplasia and prostatitis, the level of PSA will also increase, resulting in a series of negative results of prostate cancer biopsy, increasing the patient's Medical expenses and psychological burden. [0003] At present, there are urine detection kits based on PCA...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/166
Inventor 赵凯
Owner 赵凯
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