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Prostate cancer diagnosis and prognosis evaluation kit

A kit and reagent technology, applied in the biological field, can solve the problems of lack of distribution characteristics, unfavorable diagnosis and prognosis of prostate cancer, and achieve high sensitivity and specificity

Pending Publication Date: 2021-06-25
SUZHOU HONGYUAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, in situ fluorescence hybridization is used clinically to detect the instability of some regions of chromosomes, but the distribution characteristics of chromosome instability at the whole genome level of prostate cancer patients are lacking, which is not conducive to comprehensively guiding the diagnosis and prognosis of prostate cancer

Method used

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  • Prostate cancer diagnosis and prognosis evaluation kit
  • Prostate cancer diagnosis and prognosis evaluation kit
  • Prostate cancer diagnosis and prognosis evaluation kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Screening of Chromosomal Unstable Regions and Gene Variation Regions

[0057] The nucleic acid library of 106 prostate patients and 113 healthy individuals diagnosed by clinicopathology was constructed, and high-throughput sequencing of the whole genome and transcriptome or targeted capture sequencing of the target region was carried out on the computer, and the chromosome copy number and gene variation were analyzed. Identify regions of chromosomal instability and genetic variation. The differences in chromosomal instability and gene variation between tumor patients and control groups were analyzed and counted.

[0058] After the above operations, 16 common chromosomal instability regions and gene variation regions that are highly related to the prostate were found, including 2q, 3q, 5q, 6q, 7p, 7q, 8p, 8q, 9p, 12q, 13q, 14q, 16q, 18q, PCA3, ERG.

[0059] From the perspective of carrying frequency, the 16 chromosomal instability regions and gene variation r...

Embodiment 2

[0060] Example 2 A method for obtaining exosomes

[0061] The kit used in this example was purchased from SBI Company, article number: 01.SBI.EXOTC10A-1.

[0062] a. Transfer 5mL of urine to a 15mL centrifuge tube and centrifuge at 3000g for 15min to remove cells or cell debris;

[0063] b. Transfer the supernatant to a sterile tube and add 1 mL of ExoQuick reagent;

[0064] c. Invert the centrifuge tube up and down, mix well; stand overnight at 4°C or room temperature;

[0065] d. Centrifuge at 1500g for 30min, a pale yellow to white precipitate can be seen at the bottom of the tube, suck off the supernatant;

[0066] e. Centrifuge at 1500g for 5 minutes, and remove the residual supernatant;

[0067] f. Add sterile distilled water or PBS to resuspend the exosome pellet.

Embodiment 3

[0068] Example 3 Urine exfoliated cell genomic DNA and total RNA co-extracted

[0069] The kit used in this example was purchased from QIAGEN (Product No.: 80204).

[0070] a. Transfer the obtained urine sample to a 15mL centrifuge tube, centrifuge at 1600g for 10min, pour off the supernatant carefully, and then carefully discard the remaining supernatant with a pipette gun.

[0071] b. Add 350μL 1×Buffer RL / DTT to a new 1.5mL centrifuge tube, and carefully pump the lysate at least 5 times with a disposable 20-gauge needle syringe to break up the cells.

[0072] c. Put the DNA purification column in a 2mL collection tube. Transfer the cell lysate to a gDNA filter column. Centrifuge at 14000g for 2min.

[0073] d. Keep the DNA purification column and extract according to n-s steps.

[0074] e. Add an equal volume of 70% ethanol to the filtrate, pipette 3-5 times.

[0075] f. Put the RNA purification column in a 2mL collection tube. Transfer ≤750μL of the mixture to the co...

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Abstract

The invention provides a group of combination of a chromosome instability region and a gene variation region. The chromosome instability region and the gene variation region comprise 2q, 3q, 5q, 6q, 7p, 7q, 8p, 8q, 9p, 12q, 13q, 14q, 16q, 18q, PCA3 and ERG. The content of 16 chromosome unstable regions and gene variation regions in prostate cancer patients is as high as 91.5%, and when being applied to reagents or kits for screening and / or diagnosing and / or prognostically evaluating prostate cancer, the very high sensitivity and specificity and very good clinical application prospect can be achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a reagent kit for prostate cancer screening, diagnosis and prognosis evaluation. Background technique [0002] Prostate cancer (PC) refers to epithelial malignant tumors that occur in the prostate gland, and is the most common malignant tumor in the male genitourinary system. The pathological types of prostate cancer include adenocarcinoma (acinar adenocarcinoma), ductal adenocarcinoma, urothelial carcinoma, squamous cell carcinoma and adenosquamous carcinoma, of which adenocarcinoma accounts for more than 95%. Prostate cancer is usually referred to as prostate adenocarcinoma. [0003] At present, the screening and diagnosis of prostate cancer mainly rely on PSA (prostate specific antigen, prostate specific antigen), digital rectal examination and needle biopsy. Among them, PSA screening is the most widely used, but benign prostatic hyperplasia also has signs of elevate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11C40B50/06
CPCC12Q1/6886C12N15/11C40B50/06C12Q2600/156C12Q2600/118C12Q2600/166
Inventor 钱自亮王艳红王白云徐文胜
Owner SUZHOU HONGYUAN BIOTECH CO LTD
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