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Prostatic cancer diagnosis, classification or prognosis marker, detection reagent or kit, system and application thereof

A prognostic marker, prostate cancer technology, applied in the field of medical diagnosis, can solve the problems of low specificity, imperfect PSA, poor sensitivity and specificity of prostate cancer, etc., and achieve a good indicator effect.

Active Publication Date: 2019-03-22
上海晟燃生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But men at risk for occult prostate cancer despite low serum PSA levels
[0005] Although PSA is considered a good prostate cancer marker in the medical community, it is considered an imperfect marker due to the following reasons: PSA is organ specific but not cancer specific, high false positive rate and low specificity Leading to a lot of unnecessary prostate biopsies and psychological problems
PSA has poor sensitivity and specificity in the diagnosis of prostate cancer, especially when PSA is 4~10ng / mL; PSA cannot distinguish between in situ cancer and fatal distant metastatic cancer

Method used

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  • Prostatic cancer diagnosis, classification or prognosis marker, detection reagent or kit, system and application thereof
  • Prostatic cancer diagnosis, classification or prognosis marker, detection reagent or kit, system and application thereof
  • Prostatic cancer diagnosis, classification or prognosis marker, detection reagent or kit, system and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0051] This embodiment provides a method for detecting the concentration ratio of serum exosome-derived GCP2 to exosome-derived CD63, including:

[0052] 1. Establishment of the basic detection method: use the exosome extraction kit (QIAGEN: exoEasy Maxi Kit or miRCURY Exosome Kits; Thermofisher: Total Exosome Isolation Reagent fromserum) to extract the total exosomes in the serum and dissolve them in In the PBS solution, an exosome-PBS solution was formed. The GCP2 and CD63 protein concentrations in the exosome-PBS solution were detected with an ELISA kit (the ELISA kit for detecting GCP2 is Human PSMA / FOLH1 DuoSet ELISA from R&DSystem; the ELISA kit for detecting CD63 is LifespanbiosciencesHuman CD63 ELISA Kit Sandwich ELISA), wherein 100 μL exosome-PBS solution was used for the detection of GCP2, and 100 μL exosome-PBS solution was used for CD63. The ratio of GCP2 concentration to CD63 concentration was calculated.

[0053] 2. Optimize the amount of PBS solution used to d...

Embodiment 2

[0057] This embodiment provides a particle number ratio of serum GCP2 positive exosomes to CD63 positive exosomes (F exo / T exo ) detection methods, including:

[0058] 1. Establishment of the basic detection method: use an exosome extraction kit (QIAGEN: exoEasy Maxi Kit or miRCURY Exosome Kits; Thermofisher: Total Exosome Isolation Reagent fromserum) to extract the total exosomes in the serum and dissolve them in In the PBS solution, an exosome-PBS solution was formed.

[0059] Incubate CD63 antibody with magnetic beads overnight at 4°C to form CD63 magnetic bead solution. Then 200 μL of exosome-PBS solution was diluted 100 times to 20 ml and 500 μL of exosome dilution was taken from 20 ml. Add 50 μL CD63 magnetic bead solution to 500 μL exosome dilution solution and incubate at 37°C for 1 hour. Place the above-incubated mixture on a magnetic stand to precipitate the magnetic beads at the bottom of the eppendorf centrifuge tube. After aspirating the supernatant, rinse t...

Embodiment 3

[0063] The purpose of this example is to evaluate the effect of the concentration ratio of serum exosome-derived GCP2 and exosome-derived CD63 on the diagnosis of prostate cancer:

[0064] 1. Enrollment samples: 30 subjects with clinical diagnosis results and known serum PSA concentration were selected as enrollment samples, and the specific information is shown in Table 2.

[0065] 2. Detect the concentration ratio of serum exosome-derived GCP2 and serum exosome-derived CD63 in the incoming sample (see Example 1 for the specific method, dissolve the exosome extract of 1mL serum sample in 50μL PBS solution, divide into 25 μL (i.e. ratio 25 μL group), used to detect GCP2 or CD63; or, the exosome extract of 1 mL serum sample was dissolved in 200 μL PBS solution, and divided into 100 μL (i.e. ratio 100 μL group), used to detect GCP2 or CD63), And draw the ROC curve according to the known diagnostic results and the detected concentration ratio. The result of described concentrati...

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Abstract

The invention relates to a prostatic cancer diagnosis, classification and / or prognosis marker, a detection reagent or kit, a system and application thereof. The prostatic cancer diagnosis marker set disclosed by the invention is prepared from exosome-derived GCP2 and exosome-derived CD63 or GCP2 positive exosome and CD63 positive exosome. The prostatic cancer diagnosis marker set has the advantages of high flexibility and good specificity in prostatic cancer diagnosis, classification and / or prognosis prediction and has a good indication effect on prostatic cancer diagnosis, classification and / or prognosis prediction.

Description

technical field [0001] The present invention relates to the field of medical diagnosis, in particular, to prostate cancer diagnosis, grading and / or prognostic markers, detection reagents or kits, systems and applications thereof. Background technique [0002] Worldwide, the incidence of prostate cancer ranks second among male malignancies. At present, the incidence of prostate cancer in the United States has surpassed that of lung cancer, becoming the first tumor that endangers men's health. The incidence of prostate cancer in my country is low, but it has shown a significant upward trend in recent years. Since 2008, prostate cancer has become the malignant tumor with the highest incidence rate in the urinary system. The incidence rate of prostate cancer in China varies greatly between urban and rural areas, especially in large cities. In 2017, 1.22 million prostate cancer patients were diagnosed in China, nearly The five-year compound growth rate is around 12%. [0003] ...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/57434G01N33/57484G01N33/6893
Inventor 王家亮
Owner 上海晟燃生物科技有限公司
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