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FISH probe and kit for identifying peripheral blood circulation tumor cells (CTCs) of urothelial carcinoma patient

A technology for peripheral blood circulation and urothelial carcinoma, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of difficult CTCs and other problems, and achieve the effect of reducing time, accurate counting and identification

Inactive Publication Date: 2016-10-05
广州迈景基因医学科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the epithelial-mesenchymal transition of some CTCs, the expression of biomarkers may undergo complex changes, and it is difficult for the above techniques to accurately identify rare CTCs in peripheral blood.

Method used

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  • FISH probe and kit for identifying peripheral blood circulation tumor cells (CTCs) of urothelial carcinoma patient

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Probes

[0031] A FISH probe for identification of circulating tumor cells in the peripheral blood of patients with urothelial carcinoma according to an embodiment of the present invention is the centromere of chromosome 3 and the centromere of chromosome 7; wherein, the centromere of chromosome 3 carries Green fluorescent marker, chromosome 7 centromere with red fluorescent marker, and the mass ratio of chromosome 3 centromere and chromosome 7 centromere is 1:1.

Embodiment 2

[0032] Embodiment 2: Kit

[0033] A kit for identifying circulating tumor cells in peripheral blood of patients with urothelial carcinoma according to an embodiment of the present invention, which contains the probe described in Embodiment 1, fixative solution, blocking solution, hybridization buffer solution and counterstaining agent;

[0034] Among them, the fixative solution contains 2-8% paraformaldehyde and 25-75% ethanol; the blocking solution is: 0.1% Triton-X100, 3% goat serum and 3% BSA are added to PBS; the hybridization buffer solution consists of the following components : 10% dextran sulfate, 10mmol / L NaCl, 50% formamide, 0.1% sodium pyrophosphate, 0.2% polyvinylpyrroline, 0.2% FICOLL, 5mmol / L disodium EDTA, 0.1% Triton X-100, 50mmol / LTris-HCl; the counterstain is 4',6-diamidino-2-phenylindole (DAPI).

Embodiment 3

[0035] Example 3: Identification of circulating tumor cells in peripheral blood of patients with urothelial carcinoma

[0036] In this example, the kit described in Example 2 was used to identify circulating tumor cells in the peripheral blood of patients with urothelial carcinoma. The specific steps for identification are as follows:

[0037]1) Carefully observe whether the specimen is completely dry. If there is still moisture, it can be placed in an oven at 30-37°C. Observe the specimen every 10 minutes during this period. If it is dry, take it out immediately; it can also be placed in a ventilated place to ventilate the specimen. Dry completely.

[0038] 2) FISH operation

[0039] 2.1 Pre-fixation: Take 200-300 μL of fixative solution to completely cover the specimen area, fix at room temperature for 8 minutes, and pay attention not to exceed 10 minutes;

[0040] 2.2 Aging: Carefully suck off the fixative on the specimen area, put it in preheated 2×SSC (pH value 7.0) and...

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Abstract

The invention relates to a FISH probe and kit for identifying peripheral blood circulation tumor cells (CTCs) of a urothelial carcinoma patient, belonging to the field of biomedicine. The FISH probe for identifying peripheral blood CTCs of a urothelial carcinoma patient contains a chromosome 3 centromere and a chromosome 7 centromere. The kit contains the probe. By utilizing the genetic characteristics of urothelial carcinoma in combination with the diagnosis specificity of the chromosome 3 and chromosome 7, the FISH probe is utilized to label the peripheral blood enriched CTCs, thereby implementing accurate counting and identification on the urothelial carcinoma CTCs, and lowering the unnecessary waste of time, human resources and cost.

Description

technical field [0001] The invention relates to a FISH probe and a kit, in particular to a FISH probe and a kit for identifying peripheral blood circulating tumor cells of patients with urothelial carcinoma. Background technique [0002] Urothelial carcinoma mainly occurs in the renal pelvis, ureter, and bladder. The main symptom is painless hematuria. It can also manifest as symptoms of bladder irritation, hydronephrosis, and tumor metastasis. Grossly, it can be papillary, polypoid, nodular, solid, ulcerative, or diffuse transmural growth, the lesion can be solitary or multifocal, the surrounding mucosa can be normal or congested, and the microscopic appearance is carcinoma in situ Or atypical hyperplasia, gray-white cut surface, clear border but no capsule. [0003] Bladder cancer is one of the top ten malignant tumors in the world. Among the male genitourinary system tumors in my country, the morbidity and mortality of bladder cancer occupy the first place, and both are ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06
CPCC12Q1/6841C12Q2531/113C12Q2533/107C12Q2543/101
Inventor 柯尊富王连唐杨冬成
Owner 广州迈景基因医学科技有限公司
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