CN103675277A - Fluorescent probe and kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry - Google Patents
Fluorescent probe and kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry Download PDFInfo
- Publication number
- CN103675277A CN103675277A CN201310559467.5A CN201310559467A CN103675277A CN 103675277 A CN103675277 A CN 103675277A CN 201310559467 A CN201310559467 A CN 201310559467A CN 103675277 A CN103675277 A CN 103675277A
- Authority
- CN
- China
- Prior art keywords
- antibody
- rhabdomyosarcoma
- neuroblastoma
- kit
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57426—Specifically defined cancers leukemia
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
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
The present invention relates to a kind of medical treatment and detect reagent, relate in particular to a kind of flow cytometry antidiastole rhabdomyosarcoma and neuroblastoma marrow and shift and leukemic fluorescence probe and kit.
Background technology
Clinically, in children's marrow, there is tumour cell commonly leukaemia, rhabdomyosarcoma and neuroblastoma, correctly differentiate that three has important directive significance to patient's treatment.Rhabdomyosarcoma and neuroblastoma are the common malignant entity tumors of children, and children's life is had to serious threat and harm.Some patients were can obtain rehabilitation by operative treatment.But marrow easily occurs this disease shifts, and often jeopardizes infant life, need take in time the even treatment measure such as autologous stem cell transplantation of chemotherapy, radiotherapy.Therefore early detection, early diagnosis and early treatment marrow being shifted is the key that improves cure rate, reduces mortality ratio.
Mainly by Bone Marrow Cell Morphology inspection, diagnose rhabdomyosarcoma and neuroblastoma marrow to shift both at home and abroad at present, but cytomorphology is insensitive for the patient's of early stage transfer diagnosis, easily produce false negative result, and both cytomorphologies are similar with myeloblast to lymphoblast, very easily be misdiagnosed as leukaemia, simultaneously rhabdomyosarcoma also similar and more difficult discriminating of the cytomorphology in marrow to neuroblastoma.
Just because of the current various inspection methods that diagnosis and differential diagnosis rhabdomyosarcoma and neuroblastoma marrow are shifted, also cannot meet clinician's requirement, seek a kind of fast and convenient and accuracy and the high method of susceptibility and come diagnosis and differential diagnosis rhabdomyosarcoma very important with the transfer of neuroblastoma marrow and leukemic method.Flow cytometry (Flow Cytometry, FCM) is the high science and technology that development in recent years is got up, and it integrates computer technology, laser technology, fluid mechanics, cytochemistry, cellular immunology, has simultaneously and analyzes and sorting cells function.It not only can measure cell size, the proterties of internal particle, also can detect cell surface and cytoplasmic antigen, DNA in cell, rna content etc., Ke Dui colony cell carries out multi parameter analysis in unicellular level, on same cell, carry out the analysis of the co expression of 2-3 kind or plurality of antigens, detect at short notice and analyze a large amount of cells, and collect, store and deal with data, carry out multiparameter quantitative test, have the advantages that susceptibility and accuracy are high, the diagnosis that apply to leukaemia and lymphadenomatous marrow sample clinically at present more, and to the rare report of the diagnosis and differential diagnosis of rhabdomyosarcoma and neuroblastoma.Because both cells do not have single specific antigen mark at present, flow cytometry shifts antidiastole rhabdomyosarcoma and knurl neuroblastoma marrow and leukaemia key is the combination of Multiple Antibodies.
Summary of the invention
The invention provides a kind of flow cytometry antidiastole diagnosis rhabdomyosarcoma and neuroblastoma marrow and shift and leukemic fluorescence probe, utilize this fluorescence probe can obtain fast and accurately diagnostic result.
A kind of flow cytometry antidiastole rhabdomyosarcoma and neuroblastoma marrow shift and leukemic fluorescence probe, comprise four kinds of fluorescent-labeled antibody, described four kinds of fluorescent-labeled antibody are for all with fluorescently-labeled GD2 antibody, CD90 antibody, CD45 antibody and CD56 antibody, wherein, different antibody is with different fluorescence labelings.
Rhabdomyosarcoma is the modal soft tissue neoplasms of children, and neuroblastoma belongs to neuroendocrine tumour, and CD56 is the membrane glycoprotein N-CAM in an immunoglobulin like protein sample region, the cell surface expression in neuroderm source.CD90 and CD56 as adhesion molecule expression in human rhabdomyosarcoma cells and neuroblastoma, NK cell is mainly also expressed the CD56 isomeride of this 140kDa, and CD45 is the restrictive membrane protein molecule of haemocyte series, expression is comprising that except exo-erythrocytic candidate stem cell is to mature blood cell (comprising NK cell) film surface, therefore can to avoid mistaken diagnosis be NK cell tumour and leukaemia to CD90+CD56+/CD45-phenotype, and can be used as accusing of that judgement neuroendocrine tumor and rhabdomyosarcoma exist.
Neuroblastoma cell derives from the poor gangliocyte of differentiation, there is the expression of nerve adhesion molecule CD56 antigen and gangliosides GD2 on its surface, and do not express leukocyte common antigen (LCA) CD45, rhabdomyosarcoma is the mesenchymal a kind of malignant tumour that originates from striated muscle cell or break up to striated muscle cell, does not express gangliosides GD2.Same leukaemia does not express GD2 but it expresses leukocyte common antigen (LCA) CD45 and initial cell sign CD90, therefore can express to identify common tumour cell in this three class of rhabdomyosarcoma, neuroblastoma and leukaemia initial cell children marrow sample by the difference of these four kinds of antigens of polychrome Flow Cytometry Assay.Three has different immunophenotypes, that is:
Human rhabdomyosarcoma cells: GD2
-cD90
+cD56
+cD45
-;
Neuroblastoma cell: GD2
+cD90
+cD56
+cD45
-;
Acute leukemia initial cell: GD2
-cD90
+cD56
-cD45
+.
The present invention combines GD2, CD90, CD56 and tetra-kinds of fluorescent-labeled antibody of CD45 as probe, the immunophenotype (a pipe method) that uses the common polychrome flow cytometer of Hospitals at Present to detect cell gets final product antidiastole rhabdomyosarcoma and neuroblastoma marrow shifts and leukaemia, and specificity can reach 100%.
Concrete, described four kinds of fluorescent-labeled antibody are GD2-FITC antibody, CD90-PE antibody, CD45-PerCP antibody and CD56-APC antibody.
For the specificity that guarantees to detect, four kinds of fluorescent-labeled antibody are monoclonal antibody.
The present invention also provides a kind of Flow Cytometry antidiastole rhabdomyosarcoma and neuroblastoma marrow to shift and leukemic kit, comprising:
(1) probe solution, includes four kinds of fluorescent-labeled antibody, and wherein, four kinds of fluorescent-labeled antibody are for all with fluorescently-labeled GD2 antibody, CD90 antibody, CD45 antibody and CD56 antibody, and different antibody is with different fluorescence labelings;
(2) hemolytic agent;
(3) phosphate buffer;
(4) positive criteria product.
Described probe solution be take phosphate buffer as solvent.
Four kinds of described fluorescent-labeled antibody are specially GD2-FITC antibody, CD90-PE antibody, CD45-PerCP antibody and CD56-APC antibody.Wherein, CD90-PE can be purchased from U.S. company BD, article No.: 555596, CD45-PerCP can be purchased from U.S. company BD, article No.: 347464, CD56-APC can be purchased from U.S. company BD, article No.: 341025, GD2-FITC preparation method can be with reference to CANCER RESEARCH, 2988 pages of 46 phases in 1986, Highly selective recognition of human neuroblastoma cells by mouse monoclonal antibody to a cytoplasmic antigen.
The concentration ratio of described four kinds of fluorescent-labeled antibody is 1:1:1:1.
Consisting of of described hemolytic agent: 8.3% ammonium chloride and 15% paraformaldehyde.
Described probe solution comprises antiseptic, and described antiseptic is Sodium azide.
Described phosphate buffer also contains containing antiseptic, and described antiseptic is Sodium azide.
The addition of described Sodium azide is 0.05～0.1%, is preferably 0.1%.
The concentration of the phosphate buffer using in kit is 0.8～1M, is preferably 1M, the pH=7.4 of this phosphate buffer.
It is pointed out that chronic myelocytic leukemia is few because children are without chronic lymphocytic leukemia, if no special instructions, leukaemia mentioned in the application all refers to acute leukemia.
Compared with prior art, beneficial effect of the present invention is:
Fluorescence probe of the present invention can be combined by the tumor cell specific in marrow, no cross reaction, can be accurately by Flow cytometry rhabdomyosarcoma and neuroblastoma, whether marrow shifts fast, there is very high specificity (can reach 100%) and sensitivity, can distinguish acute leukemia initial cell simultaneously, simple and quick accurate, there is good potential applicability in clinical practice.
Accompanying drawing explanation
Fig. 1 is the Wright's staining result figure of the present invention to embodiment 1 Bone Marrow of Patients sample; Wherein, A is that rhabdomyosarcoma marrow shifts, and B is that neuroblastoma marrow shifts.
Fig. 2 is the Flow cytometry result figure of the present invention to embodiment 1 rhabdomyosarcoma Bone Marrow of Patients sample; Wherein, A:CD45-and CD56+ cell are established R1 door, and B:GD2-and CD90+ cell are established R2 door; C:FSC and SSC cell are established R3 door; R2 derives from R1, and R3 derives from R2.
Fig. 3 is the Flow cytometry result figure of the present invention to embodiment 1 neuroblastoma Bone Marrow of Patients sample; Wherein, A:CD45-and CD56+ cell are established R1 door, and B:GD2+ and CD90+ cell are established R2 door; C:FSC and SSC cell are established R3 door; R2 derives from R1, and R3 derives from R2.
Fig. 4 is the Wright's staining result figure of embodiment 1 acute leukemia initial cell.
Fig. 5 is the Flow cytometry result figure of the present invention to embodiment 1 acute leukemic patient marrow sample; Wherein, A:CD45+ and CD90+ cell are established R1 door, B: the cell that derives from R1 shows GD2-and CD45+; C: the cell that derives from R1 shows GD56-and CD45+.
Fig. 6 a is the flow cytometry sensitivity Detection result figure of the present invention to embodiment 1 rhabdomyosarcoma Bone Marrow of Patients sample (dilution ratio is 1:100).
Fig. 6 b is the flow cytometry sensitivity Detection result figure of the present invention to embodiment 1 rhabdomyosarcoma Bone Marrow of Patients sample (dilution ratio is 1:1000).
Fig. 6 c is the flow cytometry sensitivity Detection result figure of the present invention to embodiment 1 rhabdomyosarcoma Bone Marrow of Patients sample (dilution ratio is 1:10000).
Fig. 7 is the Wright's staining result figure of the present invention to embodiment 2 rhabdomyosarcoma Cerebrospinal Fluid in Patients samples.
Fig. 8 is the Flow cytometry result figure of the present invention to embodiment 2 rhabdomyosarcoma Cerebrospinal Fluid in Patients samples, and wherein, A:CD45-and CD56+ cell are established R1 door, and B:GD2-and CD90+ cell are established R2 door; C:FSC and SSC cell are established R3 door; R2 derives from R1, and R3 derives from R2.
Embodiment
Below in conjunction with embodiment, further explain the present invention.
Embodiment 1
GD2-FITC(provides for oneself, with neuroblastoma cell line LAN-1 immunity BALB/c mouse, getting mouse bone-marrow-derived lymphocyte and murine myeloma cell strain NS1 is fused into hybridoma and prepares monoclonal antibody GD2 flag F ITC fluorescein, concrete technical step is with reference to magazine CANCER RESEARCH, 2988 pages of 46 phases in 1986; Highly selective recognition of human neuroblastoma cells by mouse monoclonal antibody to a cytoplasmic antigen), CD90-PE(U.S. company BD, 555596), CD45-PerCP(U.S. company BD article No.:, article No.: 347464), CD56-APC(U.S. company BD, article No.: 341025).
One, kit forms:
1) probe solution (2mL * 1 bottle):
GD2-FITC:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of marked by fluorescein isothiocyanate;
CD90-PE:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of phycoerythrin mark;
CD45-PerCP:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of paramecium phyllochlorin mark;
CD56-APC:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of allophycocyanin mark;
Composition antibody concentration ratio GD2-FITC:CD90-PE:CD45-PerCP:CD56-APC is 1:1:1:1.
2) 10 * hemolytic agent (100mL * 1 bottle): 8.3% ammonium chloride, containing 15% paraformaldehyde;
3) 10 * phosphate buffer (1M, containing 1% Sodium azide for PBS, pH=7.4) (100mL * 1 bottle);
4) positive criteria product (2mL * 1 bottle): fix and preserve the cell line (5 * 10 that liquid is processed
7individual cell/mL).
Two, the application of kit
Research object: 15 routine non-leukaemia and tumour children, 21 routine rhabdomyosarcomas, 25 routine neuroblastomas and 22 routine acute myeloblastic leukemia patients' marrow sample, marrow sample is all from Medical College of Zhejiang Univ.'s attached children's hospital outpatient service and inpatient.
Operation steps: 100 μ L bone marrow fluids, add respectively each 20 μ L of monoclonal fluorescent-labeled antibody (GD2-FITC, CD90-PE, CD45-PerCP and CD56-APC), 4 ℃ of lucifuges are reacted 30 minutes, add hemolytic agent 1mL, standing 5 minutes, add PBS5mL, 500G centrifugation 5 minutes, abandon supernatant, sediment adds PBS300 μ L, mixes, and up flow type cell instrument detects 50000 cells.
Streamed image analysis strategy:
Rhabdomyosarcoma: 1, CD45-and CD56+ establish R1 door; 2, GD2-and CD90+ cell are established R2 door; 3, FSC and SSC cell are established the analysis of R3 door; 4, calculate the positive cell number in R3 door.(R2 derives from R1; R3 derives from R2).
Neuroblastoma: 1, CD45-and CD56+ establish R1 door; 2, GD2+ and CD90+ cell are established R2 door; 3, FSC and SSC cell are established the analysis of R3 door; 4, calculate the positive cell number in R3 door.(R2 derives from R1; R3 derives from R2).
Acute leukemia initial cell: 1, CD45+ and CD90+ establish R1 door; 2, analyze CD45+ and GD2-cell; 3, analyze CD45+ and CD56-cell.
Set up negative control simultaneously.
Positive findings is as follows with immunophenotype judgement cell type:
Phenotype is GD2
-cD90
+cD56
+cD45
-for human rhabdomyosarcoma cells;
Phenotype is GD2
+cD90
+cD56
+cD45
-for neuroblastoma cell;
Phenotype is GD2
-cD90
+cD56
-cD45
+for acute leukemia initial cell.
Detection sensitivity is measured: respectively human rhabdomyosarcoma cells is diluted to 1:100 with normal marrow cell, and 1:1000, the ratio of 1:10000, detects in order to upper method equally.
Statistical method: utilize software SPSS11.5 (SPSS Inc., Chicago, IL, USA), the susceptibility of assay diagnostic method and specificity.
The testing result of all types of samples as shown in Figure 1, Figure 2, shown in Fig. 3, Fig. 4, Fig. 5, Fig. 6 a, Fig. 6 b, Fig. 6 c.
Result shows: 15 routine non-leukaemia and tumour children all do not detect above three para-immunity phenotype cells; In 21 routine human rhabdomyosarcoma cells patients, 5 examples detect GD2
-cD90
+cD56
+cD45
-immunophenotype cell, clinical this 5 example of finally making a definite diagnosis is for the transfer of rhabdomyosarcoma marrow; In 25 routine neuroblastoma patients, 11 examples detect GD2
+cD90
+cD56
+cD45
-immunophenotype cell, clinical this 11 example of finally making a definite diagnosis is for the transfer of neuroblastoma marrow; 22 routine acute leukemia cellses are GD2
-cD90
+cD56
-cD45
+immunophenotyping; Flow cytometry is 100% to the specificity of three class tumour cell diagnosis in marrow, and susceptibility is 1/10
-4(in 10000 normal cells, 1 tumour cell can be detected).
Embodiment 2
One, kit forms:
1) probe solution (2mL * 1 bottle):
GD2-FITC:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of marked by fluorescein isothiocyanate;
CD90-PE:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of phycoerythrin mark;
CD45-PerCP:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of paramecium phyllochlorin mark;
CD56-APC:2mL PBS(is containing 0.1% Sodium azide) containing the 100 μ g antibody proteins (50 μ g/mL) of allophycocyanin mark;
Composition antibody concentration ratio GD2-FITC:CD90-PE:CD45-PerCP:CD56-APC is 1:1:1:1.
2) 10 * hemolytic agent (100mL * 1 bottle): 8.3% ammonium chloride, containing 15% paraformaldehyde;
3) 10 * phosphate buffer (1M, containing 1% Sodium azide for PBS, pH=7.4) (100mL * 1 bottle);
4) positive criteria product (2mL * 1 bottle): fix and preserve the cell line (5 * 10 that liquid is processed
7individual cell/mL).
Two, the application of kit
Research object: 2 examples are suspected to be that rhabdomyosarcoma cerebrospinal fluid shifts patient's samples of CSF that punctures.
Operation steps, analysis strategy and statistical method are with embodiment 1.
Result shows: 2 routine rhabdomyosarcoma clinical signs of suspected are that cerebrospinal fluid transfer patient punctures cerebrospinal fluid through the detection of this antibody combination flow cytometer, find GD2
-cD90
+cD56
+cD45
-immunophenotype cell (Fig. 7, Fig. 8), this two example of final clinical diagnosis is the transfer of rhabdomyosarcoma cerebrospinal fluid, and carries out intrathecal chemotherapy and craniospinal radiation therapy.Show that fluorescence probe of the present invention can monitor rhabdomyosarcoma cerebrospinal fluid and shift, instruct clinical rational drug use.
Claims (9)
1. a flow cytometry antidiastole rhabdomyosarcoma and neuroblastoma marrow shift and leukemic fluorescence probe, comprise four kinds of fluorescent-labeled antibody, it is characterized in that, described four kinds of fluorescent-labeled antibody are for all with fluorescently-labeled GD2 antibody, CD90 antibody, CD45 antibody and CD56 antibody, wherein, different antibody is with different fluorescence labelings.
2. fluorescence probe as claimed in claim 1, is characterized in that, described four kinds of fluorescent-labeled antibody are GD2-FITC antibody, CD90-PE antibody, CD45-PerCP antibody and CD56-APC antibody.
3. fluorescence probe as claimed in claim 1 or 2, is characterized in that, four kinds of fluorescent-labeled antibody are monoclonal antibody.
4. Flow Cytometry antidiastole rhabdomyosarcoma and neuroblastoma marrow shift and a leukemic kit, it is characterized in that, comprising:
(1) probe solution, includes four kinds of fluorescent-labeled antibody, and wherein, four kinds of fluorescent-labeled antibody are for all with fluorescently-labeled GD2 antibody, CD90 antibody, CD45 antibody and CD56 antibody, and different antibody is with different fluorescence labelings;
(2) hemolytic agent;
(3) phosphate buffer;
(4) positive criteria product.
5. kit as claimed in claim 4, is characterized in that, described probe solution be take phosphate buffer as solvent.
6. kit as claimed in claim 4, is characterized in that, four kinds of described fluorescent-labeled antibody are GD2-FITC antibody, CD90-PE antibody, CD45-PerCP antibody and CD56-APC antibody.
7. kit as claimed in claim 4, is characterized in that, the concentration ratio of four kinds of fluorescent-labeled antibody is 1:1:1:1.
8. kit as claimed in claim 4, is characterized in that, described probe solution contains antiseptic.
9. kit as claimed in claim 8, is characterized in that, described antiseptic is Sodium azide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310559467.5A CN103675277B (en) | 2013-11-12 | 2013-11-12 | Flow cytometry antidiastole rhabdomyosarcoma and neuroblastoma bone marrow neoplasms and leukemic fluorescence probe and kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310559467.5A CN103675277B (en) | 2013-11-12 | 2013-11-12 | Flow cytometry antidiastole rhabdomyosarcoma and neuroblastoma bone marrow neoplasms and leukemic fluorescence probe and kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103675277A true CN103675277A (en) | 2014-03-26 |
| CN103675277B CN103675277B (en) | 2015-10-28 |
Family
ID=50313463
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310559467.5A Expired - Fee Related CN103675277B (en) | 2013-11-12 | 2013-11-12 | Flow cytometry antidiastole rhabdomyosarcoma and neuroblastoma bone marrow neoplasms and leukemic fluorescence probe and kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103675277B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105758783A (en) * | 2016-05-05 | 2016-07-13 | 苏州大学 | Method for detecting protease-4 of human airway trypsin sample by using flow cytometry |
| CN112391263A (en) * | 2020-11-18 | 2021-02-23 | 深圳市儿童医院 | Neuroblastoma circulating tumor cell capturing chip and manufacturing method thereof |
| CN115389766A (en) * | 2022-08-29 | 2022-11-25 | 深圳市瑞格生物科技有限公司 | Marker for diagnosing whether neuroblastoma has bone marrow infiltration or not and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0530991A (en) * | 1991-07-31 | 1993-02-09 | Taisho Pharmaceut Co Ltd | Ganglioside-resistant monoclonal antibody |
| CN1284568A (en) * | 1999-08-13 | 2001-02-21 | 杨梦甦 | Disease detecting method utilizing leucocyte disserentiating antigen gene chip |
| US6306575B1 (en) * | 1995-06-16 | 2001-10-23 | Stemcell Technologies, Inc. | Methods for preparing enriched human hematopoietic cell preparations |
| CN102768283A (en) * | 2012-07-23 | 2012-11-07 | 北京大学人民医院 | Kit for detecting or supplementarily detecting medullary leukemia cell differentiation stages |
| CN103347894A (en) * | 2010-06-19 | 2013-10-09 | 纪念斯隆-凯特林癌症中心 | Anti-GD2 antibodies |
-
2013
- 2013-11-12 CN CN201310559467.5A patent/CN103675277B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0530991A (en) * | 1991-07-31 | 1993-02-09 | Taisho Pharmaceut Co Ltd | Ganglioside-resistant monoclonal antibody |
| US6306575B1 (en) * | 1995-06-16 | 2001-10-23 | Stemcell Technologies, Inc. | Methods for preparing enriched human hematopoietic cell preparations |
| CN1284568A (en) * | 1999-08-13 | 2001-02-21 | 杨梦甦 | Disease detecting method utilizing leucocyte disserentiating antigen gene chip |
| CN103347894A (en) * | 2010-06-19 | 2013-10-09 | 纪念斯隆-凯特林癌症中心 | Anti-GD2 antibodies |
| CN102768283A (en) * | 2012-07-23 | 2012-11-07 | 北京大学人民医院 | Kit for detecting or supplementarily detecting medullary leukemia cell differentiation stages |
Non-Patent Citations (4)
| Title |
|---|
| CRISTIANE S. FERREIRA-FACIO 等: "Contribution of Multiparameter Flow Cytometry Immunophenotyping to the Diagnostic Screening and Classification of Pediatric Cancer", 《PLOS ONE》 * |
| 张瑛 等: "CD56在急性白血病细胞中的表达及临床意义", 《中国实验血液学杂志》 * |
| 汤宏峰 等: "外周血和骨髓微量神经母细胞瘤细胞检测的临床意义", 《中华小儿外科杂志》 * |
| 肖平 等: "白血病干细胞相关抗原在急性白血病细胞中的表达", 《中华实用诊断与治疗杂志》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105758783A (en) * | 2016-05-05 | 2016-07-13 | 苏州大学 | Method for detecting protease-4 of human airway trypsin sample by using flow cytometry |
| CN105758783B (en) * | 2016-05-05 | 2019-07-26 | 苏州大学 | A method of utilizing Flow cytometry popularity road trypsin like proteases 4 |
| CN112391263A (en) * | 2020-11-18 | 2021-02-23 | 深圳市儿童医院 | Neuroblastoma circulating tumor cell capturing chip and manufacturing method thereof |
| CN112391263B (en) * | 2020-11-18 | 2022-05-27 | 深圳市儿童医院 | Neuroblastoma circulating tumor cell capturing chip and manufacturing method thereof |
| CN115389766A (en) * | 2022-08-29 | 2022-11-25 | 深圳市瑞格生物科技有限公司 | Marker for diagnosing whether neuroblastoma has bone marrow infiltration or not and application thereof |
| CN115389766B (en) * | 2022-08-29 | 2023-09-22 | 深圳市瑞格生物科技有限公司 | Marker for diagnosing whether neuroblastoma is subjected to bone marrow infiltration and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103675277B (en) | 2015-10-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11806714B2 (en) | Selective delivery of material to cells | |
| Gorges et al. | Heterogeneous PSMA expression on circulating tumor cells-a potential basis for stratification and monitoring of PSMA-directed therapies in prostate cancer | |
| Kirby et al. | Functional characterization of circulating tumor cells with a prostate-cancer-specific microfluidic device | |
| JP6254951B2 (en) | Method for detecting 5T4-positive circulating tumor cells in a mammalian subject and method for diagnosis of 5T4-positive cancer | |
| US11280792B2 (en) | Apparatus for detecting the degree of malignancy of each of circulating tumor cells | |
| AU2011241174B2 (en) | Method and kit for cancer diagnosis | |
| Wang et al. | A preliminary investigation of the relationship between circulating tumor cells and cancer stem cells in patients with breast cancer | |
| O'Shannessy et al. | Isolation of circulating tumor cells from multiple epithelial cancers with apostream® for detecting (or monitoring) the expression of folate receptor alpha | |
| Dowling et al. | Optimization and validation of an 8-color single-tube assay for the sensitive detection of minimal residual disease in B-cell chronic lymphocytic leukemia detected via flow cytometry | |
| Li et al. | A new score including CD43 and CD180: Increased diagnostic value for atypical chronic lymphocytic leukemia | |
| CN103675277B (en) | Flow cytometry antidiastole rhabdomyosarcoma and neuroblastoma bone marrow neoplasms and leukemic fluorescence probe and kit | |
| Bento et al. | Performance of eight‐color dry antibody reagent in the detection of minimal residual disease in chronic lymphocytic leukemia samples | |
| Wang et al. | Enrichment of circulating myeloma cells by immunomagnetic beads combined with flow cytometry for monitoring minimal residual disease and relapse in patients with multiple myeloma | |
| Zhang et al. | Enumeration and molecular characterization of circulating tumor cell using an in vivo capture system in squamous cell carcinoma of head and neck | |
| Johansson et al. | Detection of CAR‐T19 cells in peripheral blood and cerebrospinal fluid: An assay applicable to routine diagnostic laboratories | |
| CN102768283A (en) | Kit for detecting or supplementarily detecting medullary leukemia cell differentiation stages | |
| Okumura et al. | Flow cytometric detection of circulating tumor cells using a candidate stem cell marker, p75 neurotrophin receptor (p75NTR) | |
| Li et al. | Microfluidic chip for cancer cell detection and diagnosis | |
| US7615358B2 (en) | Method for determining the concentration of vital epithelial tumor cells in a body fluid | |
| JP6206953B2 (en) | Method for testing the likelihood of developing adult T-cell leukemia | |
| Vaskova et al. | Cytometric analysis of cell suspension generated by cavitron ultrasonic surgical aspirator in pediatric brain tumors | |
| Mayado et al. | Detection of circulating tumor cells in blood of pancreatic ductal adenocarcinoma patients | |
| Zhang et al. | Circulating tumor cells as potential prognostic biomarkers for early-stage pancreatic cancer: A systematic review and meta-analysis | |
| EP3602051A1 (en) | Methods of using giant cell nucleic acid characterization in cancer screening, diagnostics, treatment and recurrence | |
| Wang et al. | CD19 Chimeric Antigen Receptor T-cell Shows Activity in Patients with B-cell Lymphoma Without CD19 Expression by Immunohistochemistry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20151028 Termination date: 20181112 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |