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Fluorescent probe and kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry

A technology for neuroblastoma and rhabdomyosarcoma, which is applied in measuring devices, instruments, and individual particle analysis, can solve the problems of rare reports on the diagnosis and differential diagnosis of stroma and neuroblastoma, and achieve good clinical application prospects , highly specific effects

Inactive Publication Date: 2014-03-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can not only measure cell size and internal particle properties, but also detect cell surface and cytoplasmic antigens, intracellular DNA, RNA content, etc. It can perform multi-parameter analysis on group cells at the single-cell level, that is, on the same cell The analysis of the co-expression of 2-3 or more antigens can detect and analyze a large number of cells in a short time, and collect, store and process the data, and perform multi-parameter quantitative analysis, which has the characteristics of high sensitivity and accuracy. It is mostly used in the diagnosis of bone marrow samples of leukemia and lymphoma, but the diagnosis and differential diagnosis of rhabdomyosarcoma and neuroblastoma are rarely reported

Method used

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  • Fluorescent probe and kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry
  • Fluorescent probe and kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry
  • Fluorescent probe and kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry

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Embodiment 1

[0043] GD2-FITC (self-prepared, BALB / c mice were immunized with neuroblastoma cell line LAN-1, mouse B lymphocytes were fused with mouse myeloma cell line NS1 to form hybridoma cells to prepare monoclonal antibody GD2 and labeled For FITC fluorescein, refer to the magazine CANCER RESEARCH for specific technical steps, page 2988, Issue 46, 1986; Highly selective recognition of human neuroblastoma cells by mouse monoclonal antibody to a cytoplasmic antigen), CD90-PE (BD Company of the United States, article number: 555596), CD45 -PerCP (BD Company of the United States, article number: 347464), CD56-APC (BD Company of the United States, article number: 341025).

[0044] 1. Kit composition:

[0045] 1) Probe solution (2mL×1 bottle):

[0046] GD2-FITC: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with fluorescein isothiocyanate (50μg / mL);

[0047] CD90-PE: 2mL PBS (containing 0.1% sodium azide) containing phycoerythrin-labeled 100μg antibody pr...

Embodiment 2

[0071] 1. Kit composition:

[0072] 1) Probe solution (2mL×1 bottle):

[0073] GD2-FITC: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with fluorescein isothiocyanate (50μg / mL);

[0074] CD90-PE: 2mL PBS (containing 0.1% sodium azide) containing phycoerythrin-labeled 100μg antibody protein (50μg / mL);

[0075] CD45-PerCP: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with paramecium chlorophyll protein (50μg / mL);

[0076] CD56-APC: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with isophycocyanin (50μg / mL);

[0077] The antibody concentration ratio of the composition GD2-FITC: CD90-PE: CD45-PerCP: CD56-APC is 1:1:1:1.

[0078] 2) 10×hemolytic agent (100mL×1 bottle): 8.3% ammonium chloride, containing 15% paraformaldehyde;

[0079] 3) 10× phosphate buffer saline (PBS, pH=7.4, 1M, containing 1% sodium azide) (100mL×1 bottle);

[0080] 4) Positive standard (2mL×1 bottle): ...

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Abstract

The invention discloses a fluorescent probe and a kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry. The fluorescent probe comprises four fluorescence labeled antibodies; the four fluorescence labeled antibodies comprise a GD2 antibody, a CD90 antibody, a CD45 antibody and a CD56 antibody which take fluorescence labels; the different antibodies take the different fluorescence labels. The kit comprises a probe solution, a hemolytic agent, a phosphate buffer and a positive standard product. The fluorescent probe can be respectively and specifically combined with cells of the rhabdomyosarcoma and the neuroblastoma in bone marrow and original cells of acute leukemia; three tumor cells which are common in the child bone marrow, such as the bone marrow metastasis of the rhabdomyosarcoma and the neuroblastoma and the leukemia cells, can be rapidly and accurately identified by the flow cytometry, so as to guide clinical treatment; the application prospect is very good.

Description

technical field [0001] The invention relates to a medical detection reagent, in particular to a fluorescent probe and a kit for differential diagnosis of rhabdomyosarcoma and neuroblastoma bone marrow metastasis and leukemia by flow cytometry. Background technique [0002] Clinically, leukemia, rhabdomyosarcoma, and neuroblastoma are the most common tumor cells in children's bone marrow. Correct identification of the three has important guiding significance for the treatment of patients. Rhabdomyosarcoma and neuroblastoma are common malignant solid tumors in children, which pose a serious threat and harm to children's lives. Some patients can recover through surgical treatment. However, the disease is prone to bone marrow metastasis, which often endangers the life of the child. Treatment measures such as chemotherapy, radiotherapy and even autologous stem cell transplantation should be taken in time. Therefore, early detection, early diagnosis and early treatment of bone m...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/533
CPCG01N15/14G01N33/57407G01N33/57426G01N33/582
Inventor 舒强沈红强赵正言汤永民
Owner ZHEJIANG UNIV
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