Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of urothelium carcinoma

A technique for urothelial carcinoma and chromosomes, which is applied in the fields of biotechnology and medical diagnosis, can solve problems such as lack of distribution characteristics, and achieve the effects of high sensitivity, high accuracy and simple operation

Pending Publication Date: 2019-10-11
SUZHOU HONGYUAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is a clinical method using in situ fluorescence hybridization to detect the instability of some regions of chromosomes, but it lacks the distribution characteri

Method used

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  • Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of urothelium carcinoma
  • Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of urothelium carcinoma
  • Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of urothelium carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of library adapters

[0046] Linker sequences synthesized by third-party companies:

[0047] Linker-1 (SEQ ID NO.1):

[0048] 5'-CGTGATAGATCGGAAGAGCACACGTCTGAACTCCAGTC-3';

[0049] Linker-2 (SEQ ID NO.2):

[0050] 5'-ACACTCTTTCCCTACACGACGCTCTTCCGATCTATCACGT-3'.

[0051] The 5' end of Adapter-1 is modified by phosphorylation, and the 3' end of Adapter-2 is modified by phosphorothioation between G and T.

[0052] Use the annealing buffer (Annealing buffer) to dilute the dry powder of adapter-1 and adapter-2 to 20 μM respectively, mix the two dilutions in equal proportions, put the mixture in the PCR instrument, and run the program as shown in Table 1:

[0053] Table 1

[0054]

[0055] The product was subjected to agarose electrophoresis, and the size range of the electrophoresis band was between 80-100bp.

Embodiment 2

[0057] Materials used:

[0058] human genomic DNA

[0059] DNA fragmentation reagent (Enzymatics)

[0060] DNA end repair, adding A, adding adapter enzyme reaction system (NEB)

[0061] PCR amplification reaction system (Kapa Biosystems)

[0062] Purification magnetic beads (Beckman)

[0063] Steps:

[0064] 1. Genome Fragmentation

[0065] Take 20ng of genomic DNA, use the enzyme digestion reaction system, configure the reaction system according to Table 2, and perform the reaction according to Table 3.

[0066] For cell-free DNA, such as cfDNA in blood, it does not need to undergo genome fragmentation and can directly enter the next step.

[0067] Table 2

[0068]

[0069]

[0070] Shake to mix, centrifuge (avoid air bubbles) and run the following program on the PCR instrument:

[0071] table 3

[0072]

[0073] 2. End repair plus A

[0074] Configure the end repair plus A reaction system as shown in Table 4:

[0075] Table 4

[0076]

[0077] Shake t...

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Abstract

The invention relates to the field of biotechnology and medical diagnosis, in particular to application of chromosome instable regions to preparing an agent or kit for the diagnosis of urothelium carcinoma. There are ten chromosome instable regions 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p and 17q. Samples of clinically confirmed urothelium carcinoma patients, healthy people and non-urothelium-carcinomapatients are collected, the specific chromosome instability distribution characteristics of the urothelium carcinoma are expounded by comparing chromosome abnormity conditions of the tumor patients and the contrast through whole-genome sequencing, the ten common chromosome instable regions of the urothelium carcinoma patients are disclosed, and the total carrying rate is up to 93.4%, which has great significance in the clinical diagnosis, treatment, monitoring, prognosis and evaluation of the urothelium carcinoma and provides the scientific basis for conducting early diagnosis next and formulating an individualized treatment scheme.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and medical diagnosis, specifically, a group of chromosomal instability mutations related to urothelial carcinoma, and the use of these unstable mutations in preparing reagents or reagents for diagnosing urothelial carcinoma and evaluating prognosis application in the box. Background technique [0002] Urothelial cancer (UC) refers to malignant tumors that occur in the bladder mucosa, ureteral epithelium, and renal pelvis. It is the most common malignant tumor of the urinary system and one of the ten most common tumors in the whole body. It occupies the first place in the incidence of genitourinary system tumors in my country. The pathological types of urothelial carcinoma in the histological classification of tumors of the urinary tract system in WHO "Pathology and Genetics of Tumors of the Urinary System and Male Genital Organs" in 2004 include urothelial carcinoma of the bladder, squam...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/156
Inventor 钱自亮王白云徐文胜王艳红
Owner SUZHOU HONGYUAN BIOTECH CO LTD
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