CN101968491A - Molecular pathological typing method and kit for diffuse large B cell lymphoma and application - Google Patents
Molecular pathological typing method and kit for diffuse large B cell lymphoma and application Download PDFInfo
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Abstract
The invention discloses a molecular pathological typing method of diffuse large B cell lymphoma, which takes quantum dot markers with three different wavelengths as fluorescent probes to detect the expression of CD10, Bcl-6 and MUM1 proteins in a biological sample. The invention also discloses a kit for molecular pathological typing of the diffuse large B cell lymphoma and application thereof. The molecular pathological typing method of the diffuse large B cell lymphoma obviously improves the working efficiency, adopts a reasonable treatment scheme for the diffuse large B cell lymphoma patient and is beneficial to saving the medical expense.
Description
Technical field
The present invention relates to hemooncology and medical domain, relate in particular to diffuse large b-cell lymphoma molecule pathological typing method and kit and application.
Background technology
(diffuse large B-cell lymphoma is modal non-Hodgkin lymphoma DLBCL) to diffuse large B cell lymphoma, accounts for annual about 40% of the non-Hodgkin lymphoma of just sending out.The incidence of disease of the present non-Hodgkin lymphoma of China is about 3-4/10 ten thousand, and with the speed increment of annual 2-3%, wherein diffuse large B cell lymphoma (DLBCL) is the most common, account for annual all lymphadenomatous 50%, therefore, annual national new cases are about 2-3 ten thousand.
DLBCL has significant biology heterogeneity, and patient is remarkable to the response difference of treatment, and conventional CHOP Combination chemotherapy (ring phosphinylidyne ammonia, adriamycin, vincristine and bold and vigorous mud pine) can only make about 40% patient obtain long-term remission.Along with people-mouse mosaic anti-CD-20 monoclonal antibody (Rituximab, Rituximab) application of combined chemotherapy (R-CHOP) further improves the clinical efficacy of DLBCL, but still has the patient of 30-40% to recur rapidly failing to respond to any medical treatment or treating the back, progression of disease is fast, poor prognosis.Therefore, strengthen the DLBCL biological Characteristics Study, formulate the research focus that the corresponding treatment scheme becomes present DLBCL.Currently clinical DLBCL being carried out risk stratification, mainly is that (International Prognostic Index IPI), comprising: at the age, performance status, clinical Ann Arbor save outer focus number, the lactic dehydrogenase enzyme level by stages by international prognostic index.Because this evaluation is the combination of clinical parameter, can not reflect molecular biology heterogeneity inherent in the tumour generating process, therefore can not accurately estimate for the prognosis of quite a few case, some 5 years survival rates of patient that are in low danger, low middle danger group still only have 32%.
Being applied as of biochip technology illustrated the potential biology heterogeneity of DLBCL provides a high flux platform, by the DLBCL gene expression spectrum analysis, three kinds of DLBCL hypotypes have been identified, prompting tumprigenicity B cell is in different differential periods: a kind of B of centrum germinativum cell sample DLBCL (germinal center B cell-like DLBCL that is called, GCB), this hypotype because of with identical the gaining the name of genetic marker of the normal B of centrum germinativum cellular expression; Another kind of hypotype is called activating B cell sample DLBCL, and (activated B cell-like DLBCL, ABC), the genotype of this subtype expression stimulates the back peripheral blood B cell similar to mitogenesis, and thousands of gene expression differences are arranged between GCB and the ABC.Except that above amphitypy, still have the DLBCL of 17%-40% to come somatotype with origin of cell, be collectively referred to as 3 types (type3), ABC and type3 are referred to as non-GCB (non-GCB).The DLBCL hypotype is the prognostic evaluation index that is independent of IPI, each hypotype has nothing in common with each other to the reactivity and the curative effect of traditional combined chemotherapy CHOP scheme, the five year survival rate of ABC and GCB is respectively 31% and 59%, the survival rate of GCB is significantly better than ABC, and 3 types itself have heterogeneity, do not have the survival rate of determining, but its overall prognosis situation is similar to the ABC hypotype.
Fill the air the analysis of large B cell lymphoid tumor gene chip expression spectral by 20 examples, detect some specific genes relevant with filling the air large b-cell lymphoma molecule pathological typing and prognosis, as: Bcl-2, Bcl-6, CCND2, MYC, SCYA3, FN1, FOXP1, PKC-β, HGAL, CD10 and MUM1 or the like.Therefrom filter out Three Represents and the gene of B cell differentiation different phase: CD10, Bcl-6 and MUM1, wherein CD10 and Bcl-6 are the B of centrum germinativum cell sign, and MUM1 is the back B of a centrum germinativum cell sign.
Biochip technology has been deepened the understanding to the DLBCL biological characteristics, but that the having relatively high expectations of sample (fresh specimens, suitable freezing preservation etc.), complicated operation and genetic chip are detected cost is higher because of it, so restricted its clinical practice.Application number is that 200710173600.8 application for a patent for invention is to utilize common immunohistochemistry (immunohistochemistry, IHC) method, the expression that the DLBCL sample is carried out CD10, Bcl-6 and MUM1 albumen detects, and carries out the molecular pathology somatotype by interpretation of result to filling the air large B cell lymphoid tumor.But, the method of this routine immunization groupization is to carry out the result by the means of colour developing to judge basically, the detection of three kinds of indexs just means three sections of the minimum needs of same sample, and immunohistochemical experiment is carried out in every section one time, and the result to three sections carries out interpretation respectively then.There are a lot of repeating steps in whole process, and complex operation, efficient are low.
Summary of the invention
Complex operation, inefficient technical matters when the present invention will solve common SABC method diffuse large B cell lymphoma is carried out the molecular pathology somatotype, a kind of diffuse large b-cell lymphoma molecule pathological typing method in conjunction with technology of quantum dots is provided, has significantly improved the work efficiency of experiment.
In addition, also need to provide a kind of kit and application thereof of diffuse large b-cell lymphoma molecule pathological typing.
In order to solve the problems of the technologies described above, the present invention is achieved through the following technical solutions:
In one aspect of the invention, provide a kind of diffuse large b-cell lymphoma molecule pathological typing method, may further comprise the steps:
With the quantum dot-labeled thing of three kinds of different wave lengths as fluorescence probe, the expression of CD10, Bcl-6 and MUM1 albumen in the detection of biological sample;
Carry out the molecular pathology somatotype according to testing result, if CD10 (+) or CD10 (-)/Bcl-6 (+)/MUM1 (-) show that then this diffuse large B cell lymphoma is the GCB type; If CD10 (-)/Bcl-6 (-) or CD10 (-)/Bcl-6 (+)/MUM1 (+) show that then this diffuse large B cell lymphoma is non-GCB type.
The quantum dot-labeled thing of described three kinds of different wave lengths is used for mark biological sample CD10, Bcl-6 and MUM1 albumen respectively, makes and can detect these three kinds of albumen simultaneously in same section.
Preferably, described detection realizes by the method for immunohistochemistry or organization chip.
Preferably, three kinds of monoclonal antibodies that immunohistochemistry is used, it comprises two kinds of different genera sources at least, promptly when selecting these three kinds of monoclonal antibodies, at least select for use the monoclonal antibody in two kinds of kind sources to make up, for example two kinds of mouse sources of three kinds of monoclonal antibodies, a kind of rabbit source.
In another aspect of this invention, also provide a kind of kit that is used for diffuse large b-cell lymphoma molecule pathological typing, described kit comprises:
Specific antibody and quantum dot-labeled thing at CD10 albumen;
Specific antibody and quantum dot-labeled thing at Bcl-6 albumen;
Specific antibody and quantum dot-labeled thing at MUM1 albumen.
Preferably, described quantum dot at CD10 albumen, Bcl-6 albumen and MUM1 albumen has different wavelength, is suitable for detecting simultaneously in same section these three kinds of albumen.
Described kit adopts the method for immunohistochemistry or organization chip that biological sample is detected.
In another aspect of this invention, also provide a kind of application of mentioned reagent box, be used in reference to the guiding doctor and give birth to the diffuse large B cell lymphoma patient is selected the therapeutic scheme that suits,
When the testing result of kit is CD10 (+) or CD10 (-)/Bcl-6 (+)/MUM1 (-), show that this diffuse large B cell lymphoma is the GCB type, should adopt the treatment of CHOP scheme to the diffuse large B cell lymphoma patient;
When the testing result of kit is CD10 (-)/Bcl-6 (-) or CD10 (-)/Bcl-6 (+)/MUM1 (+), show that this diffuse large B cell lymphoma is non-GCB type, should adopt the treatment of R-CHOP scheme to the diffuse large B cell lymphoma patient.
Taking the selection of above-mentioned therapeutic scheme, is that GCB type patient prognosis obviously is better than non-GCB type patient because find that in former studies the diffuse large B cell lymphoma patient adopts traditional C HOP scheme; Adopt the treatment of R-CHOP scheme, GCB type patient and non-GCB type patient prognosis no significant difference; GCB type patient adopts the treatment of CHOP scheme and adopts R-CHOP scheme treatment prognosis no significant difference.Therefore, adopt traditional CHOP scheme, GCB type patient prognosis is better than non-GCB type patient; Rituximab (Rituximab) combined chemotherapy (R-CHOP scheme) can make non-GCB type patient be benefited, and can not improve GCB type patient's curative effect.
Each course of therapy expense of Rituximab is ten thousand yuan of 10-15, is heavy financial burden for domestic most of patient.After utilizing kit of the present invention that the diffuse large B cell lymphoma patient is carried out the molecular pathology somatotype, select the proper treatment scheme, the medical expense that helps saving GCB type patient.
In another aspect of this invention, also provide a kind of application of mentioned reagent box, be used for the diffuse large B cell lymphoma patient is made prognostic evaluation.
Diffuse large b-cell lymphoma molecule pathological typing method of the present invention has following beneficial effect:
(1) the present invention is applied to the quantum dot of three kinds of emitting at different wavelengths light in the SABC detection of CD10, BCL6 and MUM1, carries out immunohistochemical experiment in a section, finishes the detection of three kinds of indexs simultaneously, has significantly improved the work efficiency of experiment.
(2) the invention solves biochip technology in carrying out DLBCL somatotype process to sample requirement height, complicated operation, problem that cost is high, have the good clinical practicality.Use the testing cost of genetic chip to be each patient 7000-9000 unit, be that each patient 200-300 unit, expense obviously reduce, and detect similar, the good reproducibility of effect and use DLBCL molecular pathology classification diagnosis kit to detect.
(3) each course of therapy expense of Rituximab is ten thousand yuan of 10-15, for domestic most of patient is heavy financial burden, because Rituximab can not improve GCB type patient's curative effect, so after identifying patient's molecular pathology somatotype, select the proper treatment scheme, the medical expense that helps saving this part patient of GCB type.
Description of drawings
The present invention is further detailed explanation below in conjunction with the drawings and specific embodiments.
Fig. 1 is a DLBCL SABC molecule parting tree synoptic diagram of the present invention;
Fig. 2 is DLBCL quantum dot molecule parting figure of the present invention.
Embodiment
In the following example, the experimental technique of unreceipted actual conditions, usually condition routinely, as " fine works molecular biology experiment guide " (F.M. Ao Sibai, R.E. James Kingston, chief editors such as J.G. Sai Deman, Ma Xuejun, the Su Yuelong translates. Beijing: Science Press, 2004) described in method carry out.
The present invention is the diffuse large b-cell lymphoma molecule pathological typing in conjunction with technology of quantum dots, a kind of quantum dot immune group detection method of diffuse large b-cell lymphoma molecule pathological typing is provided, its principle is: the quantum dot of different sizes can be sent the light of different colours by the optical excitation of single wavelength, and the photochemical stability height is suitable for use as fluorescence probe.The present invention adopts the quantum dot (can send the light of three kinds of different wave lengths during the optical excitation of same wavelength) of three kinds of different-grain diameter sizes, CD10, Bcl-6 and MUM1 albumen in the difference mark biological sample, common this mark is an indirect labelling, combine with specific antibody as quantum dot-labeled thing, CD10, Bcl-6 or MUM1 antigen protein combine in specific antibody and the biological sample.Indirect labelling also can be that quantum dot-labeled thing and two resistive connections close, two anti-combine with monoclonal antibody (special resists), CD10, Bcl-6 or MUM1 antigen protein combine in monoclonal antibody and the biological sample, promptly form " antigen-monoclonal antibody-two resists-quantum dot-labeled thing ", in this case, three kinds of used monoclonal antibodies select for use the monoclonal antibody in two kinds of kind sources to make up at least.
In embodiments of the present invention, three indexs at the needs detection, it is distributed in cell membrane (CD10) respectively, nucleus (BCL6, MUM1) in, therefore select the monoclonal antibody (CD10 in two kinds of mouse sources, MUM1) monoclonal antibody (BCL-6) with the rabbit source makes up, with quantum dot-labeled Streptavidin compound (QDs-SA) as quantum dot-labeled thing, this QDs-SA can combine with biotinylation two is anti-, when special one anti-with after respective target antigen combines, biotinylation two resists and a special resistive connection closes, and forms " antigen-special one resists-biotinylation two anti--QDs-SA " at last.The present invention adopts quantum dot-labeled thing as fluorescence probe, can in same section, detect the expression of CD10, Bcl-6 and three kinds of albumen of MUM1 simultaneously, compare with common SABC method, efficient obviously improves, and helps in time providing the proper treatment scheme for the diffuse large B cell lymphoma patient.
Embodiment 1 diffuse large B cell lymphoma quantum dot immune group detection
1. specimen collection
The DLBCL sample is all fixed with 10% neutral formalin, conventional dehydration, specimens paraffin embedding slices, hematoxylin eosin staining, by the doctor who specializes in lymthoma research by new WHO classification diagnosis.
2. sample disposal
With in baking box the dry paraffin section of crossing be dipped in the dimethylbenzene secondary 15 minutes.Take out section and place 100% absolute ethyl alcohol 10 minutes twice; Inserted 90%-70% alcohol at different levels successively each 10 minutes.Taking-up places distilled water.Take out the section in the distilled water, get rid of and dry section and go up tissue liquid on every side, lie against in the wet box, drip peroxidase blocking agent-3% methyl alcohol-H
2O
2Hatched 20 minutes in the tissue lucifuge.Distilled water flushing is inserted section in the TBS damping fluid again, soaks three times 5 minutes.Take out section, get rid of and dry tissue liquid on every side, lie against in the wet box.
Medical microwave stove antigen retrieval 20 minutes was taken out the back natural cooling 30 minutes, antigen retrieval liquid select for use Tris/EDTA (50mMTris, 2mM EDTA, PH9.0).
1% bovine serum albumin(BSA) (BSA) room temperature sealing 20 minutes.
3. quantum dot immune histochemistry is detected
At three indexs that needs detect, it is distributed in respectively in cell membrane (CD10), the nucleus (BCL6, MUM1), therefore selects the monoclonal antibody (CD10, MUM1) in two kinds of mouse sources and the monoclonal antibody (BCL-6) in rabbit source to make up.
Dropping is hatched the 2hr commentaries on classics and is spent the night for 4 ℃ with one anti-(CD10 and BCL6 mixed liquor) of antibody diluent dilution, 37 ℃ of wet boxes;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 10min;
The biotinylation goat anti-rabbit igg that dropping is diluted with antibody diluent, 37 ℃ of wet boxes are hatched 30min;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 20min;
Dropping with antibody diluent dilution by quantum dot (quantum dots, QDs) the Streptavidin compound (QDs-SA of mark, 545nm) and by two of quantum dot (QDs) mark resist (QDs-G/M IgG, 605nm is available from an ancient woman's ornament source, Wuhan technology of quantum dots Development Co., Ltd) 37 ℃ hatch 1hr;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 10min;
The MUM1 that dropping is diluted with reagent C, 37 ℃ of wet boxes are hatched 2hr;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 10min;
The anti-mouse IgG of biotinylation horse that dropping is diluted with antibody diluent, 37 ℃ of wet boxes are hatched 30min;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 20min;
The quantum dot-labeled Streptavidin compound that dropping is diluted with antibody diluent (QDs-SA, 705nm) 37 ℃ of wet boxes are hatched 30min;
The TBS-T washing (3 * 5min), TBS washing (2 * 5min); Buffering glycerine mounting, ultraviolet excitation is observed imaging under the fluorescent microscope.
4. the result judges and analyzes
The CD10 positive is positioned cell membrane, and Bcl-6, the MUM1 positive are positioned nucleus, greater than the painted positive that is judged to of 30% tumour cell.
Embodiment 1 pathological section quantum dot testing result is: 50 examples are filled the air in the large B cell lymphoid tumor sample, and the positive rate of CD10, Bcl-6, MUM1 is respectively 34%, 34%, 56%.
5. fill the air large B cell lymphoid tumor molecule parting (SABC method) and pathological section molecule parting result
CD10, Bcl-6 are as the mark in centrum germinativum cell source, and CD10 (+) or CD10 (-)/Bcl-6 (+)/MUM1 (-) is the GCB class; CD10 (-)/Bcl-6 (-) is non-GCB class.MUM1 is as the mark in back centrum germinativum cell source, if CD10 (-)/Bcl-6 (+), then classify with MUM1: MUM1 (+) is non-GCB class, MUM1 (-) is a GCB class (see figure 1), be that CD10 (-)/Bcl-6 (+)/MUM1 (+) is non-GCB class, CD10 (-)/Bcl-6 (+)/MUM1 (-) is the GCB class.
In brief, as CD10 (+) or CD10 (-)/Bcl-6 (+)/MUM1 (-), show that then this fills the air large B cell lymphoid tumor is the GCB class; As CD10 (-)/Bcl-6 (-) or CD10 (-)/Bcl-6 (+)/MUM1 (+), show that then this fills the air large B cell lymphoid tumor is non-GCB class.
DLBCL quantum dot molecule parting image as shown in Figure 2.
Embodiment 1 pathological section molecule parting result is as follows:
50 total examples are filled the air in the large B cell lymphoid tumor sample GCB18 example (36%), non-GCB32 example (64%).In 18 routine GCB, 17 examples (94.4%) CD10 (+), 1 example (5.6%) CD10 (-)/Bcl-6 (+)/MUM1 (-).In 32 routine non-GCB, 12 examples (27.5%) CD10 (-)/Bcl-6 (-)/MUM1 (+), 7 examples (21.9%) CD10 (-)/Bcl-6 (+)/MUM1 (+), 13 examples (40.6%) CD10 (-)/Bcl-6 (-)/MUM1 (-).
Embodiment 2 organization chip diffuse large B cell lymphoma quantum dot immune group detections
1. sample disposal
Preceding organization chip is placed in 60 constant temperature ovens of dewaxing toasted 30 minutes; Place dimethylbenzene I to soak 10 minutes, in dimethylbenzene II, soaked 10 minutes, in dimethylbenzene III, soaked 10 minutes, in dimethylbenzene IV, soaked 10 minutes; Soaked 5 minutes among the absolute ethyl alcohol I, in absolute ethyl alcohol II, soaked again 5 minutes; Soaked 5 minutes in 95% ethanol; Soaked 5 minutes in 85% ethanol; Soaked 5 minutes in 70% ethanol; Soaked 5 minutes in the distilled water.
The antigen hot repair is multiple:
Get a certain amount of 0.01M citrate buffer pH6.0 big fire in pressure cooker and be heated to boiling, the organization chip of dewaxing after the aquation placed on the high-temperature resistance plastice section frame put into the damping fluid that has seethed with excitement, covering pot cover buckles pressure valve and continues to be heated to jet, pressure cooker is left thermal source after the jet 1-2 of picking up counting minute and be cooled to room temperature, take out slide and washed 5 minutes with PBS with after twice of the distillation washing earlier.
2. quantum dot immune histochemistry is detected
At three indexs that needs detect, it is distributed in respectively in cell membrane (CD10), the nucleus (BCL6, MUM1), therefore selects the monoclonal antibody (CD10, MUM1) in two kinds of mouse sources and the monoclonal antibody (BCL-6) in rabbit source to make up.
Dropping is hatched the 2hr commentaries on classics and is spent the night for 4 ℃ with one anti-(CD10 and BCL6 mixed liquor) of antibody diluent dilution, 37 ℃ of wet boxes;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 10min;
The biotinylation goat anti-rabbit igg that dropping is diluted with antibody diluent, 37 ℃ of wet boxes are hatched 30min;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 20min;
Dropping with quantum dot (QDs) the labelled streptavidin compound of antibody diluent dilution (QDs-SA, 545nm) and quantum dot (QDs) mark two resist that (QDs-G/M IgG 605nm) is hatched 1hr for 37 ℃;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 10min;
The MUM1 that dropping is diluted with reagent C, 37 ℃ of wet boxes are hatched 2hr;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 10min;
The anti-mouse IgG of biotinylation horse that dropping is diluted with antibody diluent, 37 ℃ of wet boxes are hatched 30min;
The TBS-T washing (3 * 5min), drip the sealing damping fluid, 37 ℃ of wet boxes are hatched sealing 20min;
The quantum dot-labeled Streptavidin compound that dropping is diluted with antibody diluent (QDs-SA, 705nm) 37 ℃ of wet boxes are hatched 30min;
The TBS-T washing (3 * 5min), TBS washing (2 * 5min); Buffering glycerine mounting, ultraviolet excitation is observed imaging under the fluorescent microscope.
3. the result judges and analyzes
The CD10 positive is positioned cell membrane, and Bcl-6, the MUM1 positive are positioned nucleus, greater than the painted positive that is judged to of 30% tumour cell.
Embodiment 2 organization chip quantum dot testing results are:
Amount to 30 examples on the organization chip and fill the air the large B cell lymphoid tumor sample, wherein 1 example is fallen sheet, and the positive rate of CD10 (15), Bcl-6 (8), MUM1 (15) is respectively 51.7%, 27.6%, 51.7%.
4. organization chip molecule parting result
30 total examples are filled the air in the large B cell lymphoid tumor sample GCB17 example (58.6%), non-GCB12 example (41.4%).In 17 routine GCB, 15 examples (88.2%) CD10 (+), 2 examples (11.8%) CD10 (-)/Bcl-6 (+)/MUM1 (-).In 12 routine non-GCB, 8 examples (66.7%) CD10 (-)/Bcl-6 (-)/MUM1 (+), 4 examples (33.3%) CD10 (-)/Bcl-6 (-)/MUM1 (-).
The above embodiment has only expressed embodiments of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to claim of the present invention.Should be pointed out that for the person of ordinary skill of the art without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (8)
1. a diffuse large b-cell lymphoma molecule pathological typing method is characterized in that, may further comprise the steps:
With the quantum dot-labeled thing of three kinds of different wave lengths as fluorescence probe, the expression of CD10, Bcl-6 and MUM1 albumen in the detection of biological sample;
Carry out the molecular pathology somatotype according to testing result, if CD10 (+) or CD10 (-)/Bcl-6 (+)/MUM1 (-) show that then this diffuse large B cell lymphoma is the GCB type; If CD10 (-)/Bcl-6 (-) or CD10 (-)/Bcl-6 (+)/MUM1 (+) show that then this diffuse large B cell lymphoma is non-GCB type.
2. method according to claim 1 is characterized in that, described detection realizes by the method for immunohistochemistry or organization chip.
3. method according to claim 2 is characterized in that, three kinds of monoclonal antibodies that immunohistochemistry is used, and it comprises two kinds of different genera sources at least.
4. a kit that is used for diffuse large b-cell lymphoma molecule pathological typing is characterized in that, described kit comprises:
Specific antibody and quantum dot-labeled thing at CD10 albumen;
Specific antibody and quantum dot-labeled thing at Bcl-6 albumen;
Specific antibody and quantum dot-labeled thing at MUM1 albumen.
5. kit according to claim 4 is characterized in that, described quantum dot at CD10 albumen, Bcl-6 albumen and MUM1 albumen has different wavelength, is suitable for detecting simultaneously in same section these three kinds of albumen.
6. according to claim 4 or 5 described kits, it is characterized in that described kit adopts the method for immunohistochemistry or organization chip that biological sample is detected.
7. the application of the described kit of claim 4 is characterized in that, be used in reference to the guiding doctor and give birth to the diffuse large B cell lymphoma patient is selected the therapeutic scheme that suits,
When CD10 (+) or CD10 (-)/Bcl-6 (+)/MUM1 (-), should adopt the treatment of CHOP scheme to the diffuse large B cell lymphoma patient;
When CD10 (-)/Bcl-6 (-) or CD10 (-)/Bcl-6 (+)/MUM1 (+), should adopt the treatment of R-CHOP scheme to the diffuse large B cell lymphoma patient.
8. the application of the described kit of claim 4 is characterized in that, is used for the diffuse large B cell lymphoma patient is made prognostic evaluation.
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