CN109884313A - The detection kit of B-lineage Acute Lymphocyte Leukemia minimal residual - Google Patents
The detection kit of B-lineage Acute Lymphocyte Leukemia minimal residual Download PDFInfo
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Abstract
The present invention relates to technical field of medical detection, and in particular to the detection kit of B-lineage Acute Lymphocyte Leukemia minimal residual, the kit include the antibody of different fluorescein-labeled anti-CD10, CD19, CD20, CD34, CD38 and CD45.Detection kit provided by the invention includes the monoclonal antibody cocktail of 6 kinds of leukocyte differentiation antigens, in conjunction with fluorescent marker and FCM analysis technology, the abnormal cell level and its parting of minimal residual of B-lineage Acute Lymphocyte Leukemia can rapidly and accurately be identified by realizing a FCM analysis, obtained Testing index can be used as intermediate result, the prognosis situation for more intuitively judging B-lineage Acute Lymphocyte Leukemia patient has important directive significance for the formulation of leukaemic's clinical treatment.
Description
Technical field
The present invention relates to technical field of medical detection, and in particular to a kind of B-lineage Acute Lymphocyte Leukemia minimal residual
Detection kit.
Background technique
Acute lymphoblastic leukemia (acute lymphoblastic leukemia, ALL) is the leaching of early differentiation stage
Bar malignant clonal expansion disease, while it is also one group of height heterogeneity disease, wherein acute B lymphocyte is white
Blood disease (B-ALL) disease incidence accounts for about the 85% of acute lymphoblastic leukemia, seriously threatens the health of people, therefore by
Pay attention to extensive.Currently, leukaemia complete remission rate greatly improves by combined chemotherapy, but leukemia relapse is current white blood
The main problem of disease treatment, the root of recurrence is mainly from internal remaining leukaemia cell.In general, by this leukaemia through controlling
It treats the internal state for still remaining a small amount of leukaemia cell after alleviating and is defined as minimal residual disease (minimal residual
Disease, MRD).Minimal Residual Disease In Acute Lymphoblastic Leukemia level carries out dynamic prison in vivo after to Leukemia Patients alleviating
It surveys, individuation Easing approach strategy is taken according to minimal residual level, the complete remission rate of patient can be significantly improved.
Method currently used for Minimal Residual Disease In Acute Lymphoblastic Leukemia detection mainly has cytomorphology technology, thin
Born of the same parents' genetics technology, fluorescence in situ hybridization technique, Polymerase Chain Reaction and low cytometric analysis.Based on leukaemia correlation
The multi-parameter Flow Cytometry (FCM) of immunophenotype detects minimal residual, has been demonstrated the assessment that can be used for prognosis, has fast
Fast easy feature.The key point that FCM detects remaining leukaemia cell is normal differentiation precursor to be identified and abnormal white blood
Sick cell, merely with a kind of single leukocyte differentiation antigen as detection minimal residual disease specificity immunological marker more
Therefore difficulty mostly uses the group of the antigen in acute lymphoblastic leukemia cell and normal marrow cell differential expression at present
It closes, carries out the parting classification of cell.However, it is micro- still to lack the immunological marker object progress with high susceptibility and specificity at present
The detection of the efficiently and accurately of small residual level, moreover, there has been no fixation refers in combination about fixed index or single index is used
Research in terms of mark assessment patient's prognosis and associated treatment effect.
Currently, the judgment criteria of minimal residual detection kit used in the prior art is more single, it is anxious for detection
Property bone-marrow-derived lymphocyte leukaemia minimal residual accuracy it is poor, omission factor is higher, therefore, need exploitation have high accuracy
With the B-lineage Acute Lymphocyte Leukemia minimal residual detection reagent of recall rate.
Summary of the invention
To solve the technical problems existing in the prior art, the object of the present invention is to provide a kind of acute B lymphocyte is white
The detection kit of blood disease minimal residual.
Inventor by the expression of the leukocyte differentiation antigen to 1036 B-lineage Acute Lymphocyte Leukemia patients into
Row tests and analyzes, and is compared with the leucocyte phenotype of normal person, it was found that B-lineage Acute Lymphocyte Leukemia patient's is white thin
The immunophenotype of the generally existing regularity of born of the same parents' differentiation antigen, and abnormal immune phenotype disclosed in the prior art is combined, through excessive
Amount differentiation antigen detection antibody combination screening, creatively find, using CD10 antibody, CD19 antibody, CD20 antibody,
The combination of CD34 antibody, CD38 antibody and CD45 antibody carries out the detection of the minimal residual of B-lineage Acute Lymphocyte Leukemia, has
Higher recall rate and accuracy；On the basis of determining above-mentioned detection antibody combination, by largely screening and optimizing, the present invention
The evaluation index of detection has been determined.For the B-ALL of any immunophenotype exception, there may be CD34+CD10+CD19+CD38+CD20-With CD34-CD10+CD19+CD38+CD20+The cell of two kinds of phenotypes, the present invention by a large amount of data statistics, in conjunction with facing
Bed data, creatively discovery uses CD34+CD10+CD19+CD38+CD20-With CD34-CD10+CD19+CD38+CD20+Two kinds
The sum of ratio of phenotype cells carries out the evaluation of minimal residual and prognosis as fixed index, single with using in the prior art
The on-fixed index of abnormal immune phenotype is compared, and has higher recall rate and accuracy.Phenotype is CD34+CD10+CD19+
CD38+CD20-Cell proportion and phenotype be CD34-CD10+CD19+CD38+CD20+Cell proportion and≤1.5 patient,
Its recurrence rate is significantly higher than the patient (P < 0.05) of its value > 1.5, i.e., as the sum of above-mentioned two fixed index > 1.5, prognosis
Preferably, long-term dynamics monitoring These parameters can significantly improve the recall rate of patient's minimal residual disease.
Firstly, the present invention provides a kind of detection kit of B-lineage Acute Lymphocyte Leukemia minimal residual, the kit
Including CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody.
For convenient for detection, the present invention in, the CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody
Contain different fluorescein labels with CD45 antibody.
Kit of the present invention includes six kinds of fluorescein-labeled antibody of difference, and inventor has found that various antibody use
Different fluorescein labels, the specificity of detection is different with susceptibility,
Preferably, in the present invention, the CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and
The fluorescein label of CD45 antibody is respectively any one in PE, APC, APC-cy7, PerCP, V450, V500.
It is largely combined and screening experiment, inventor has determined six kinds of higher fluoresceins of detection efficiency, and is final true
The standard to leukaemia minimal residual cell once can be realized using flow cytometer for the combination for having determined fluorescein and antibody
Really, fast typing.Therefore, as the preferred embodiment of the present invention, the CD10 antibody, CD19 antibody, CD20 antibody, CD34
Antibody, CD38 antibody and CD45 antibody according to sequence from front to back successively respectively by PE, APC, APC-cy7, PerCP, V450,
V500 fluorescein label.
The i.e. described kit include CD10-PE, CD19-APC, CD20-APC-cy7, CD34-PerCP, CD38-V450 and
CD45-V500。
In the present invention, the anti-CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 are anti-
Body is monoclonal antibody.
Preferably, CD10 antibody of the present invention, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45
Antibody is the monoclonal antibody in mouse or rabbit source.
Detection kit of the present invention further includes erythrocyte cracked liquid, antibody dilution reagent and phosphate buffer
One of (PBS) or it is a variety of.
Wherein, the antibody dilution reagent includes following component: phosphate buffer and fetal calf serum.
Preferably, the antibody dilution reagent includes following component: fetal calf serum 1%~2%；Phosphate buffer 0.01
~0.05M, pH are 7.2~7.4.
The phosphate buffer is the phosphate buffer of sterilized (such as high temperature).
Inventor it has furthermore been found that carry out minimal residual FCM analysis when, using specific amount ratio
CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody, the recall rate of detection and accuracy into
One step improves.
In the present invention, the CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody exist
Mass ratio when for minimal residual detection is any one or more following:
(1) mass ratio of the antibody of CD10, CD20 and CD38 is 2:5:20~1:5:5；
(2) mass ratio of the antibody of CD34 and CD19 is 3:1~1:2；
(3) mass ratio of the antibody of CD45 and CD19 is 4:1~8:1.
Preferably, the CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody with
Mass ratio when minimal residual detection is any one or more following:
(1) mass ratio of CD10 antibody, CD20 antibody and CD38 antibody is 2:5:20~1:5:8；
(2) mass ratio of CD34 antibody and CD19 antibody is 3:1~2:1；
(3) mass ratio of CD45 antibody and CD19 antibody is 6:1~8:1.
As the preferred embodiment of the present invention, the CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38
The mass ratio of antibody and CD45 antibody when detecting for minimal residual is 2:5:5:13:20:40.
Kit of the present invention is to be detected using stream type cell analyzer when for minimal residual detection, root
According to obtained various fluorescein-labeled fluorescence signal intensities, one in CD10, CD19, CD20, CD34, CD38, CD45 is determined
Kind or a variety of cell proportions for being expressed as positive or negative, judge minimal residual situation.
Specifically, working procedure of the kit when detecting for minimal residual includes the following steps:
(1) sample is acquired, sample is pre-processed；
(2) it is anti-that the fluorescein-labeled CD10 antibody, CD19 antibody, CD20 antibody, CD34 are added in the sample
Body, CD38 antibody and CD45 antibody carry out the fluorescence antibody label of sample；
(3) after carrying out red blood cell dissolution process to the sample, using flow cytomery；
(4) according to flow cytomery as a result, determine one of CD10, CD19, CD20, CD34, CD38, CD45 or
A variety of cell proportions for being expressed as positive or negative, judge minimal residual situation.
Preferably, the sample includes being selected from one of marrow, blood.
In the present invention, the judgment method of the minimal residual situation is selected from the one or more of following method:
(1) detection phenotype is CD34 respectively+CD10+CD19+CD38+CD20-Cell proportion and phenotype be CD34-CD10+
CD19+CD38+CD20+Cell proportion, according to phenotype be CD34+CD10+CD19+CD38+CD20-Cell proportion be with phenotype
CD34-CD10+CD19+CD38+CD20+The outer minimal residual of the sum of cell proportion judgement and its prognosis situation；
Preferably, when phenotype is CD34+CD10+CD19+CD38+CD20-Cell proportion and phenotype be CD34-CD10+
CD19+CD38+CD20+The sum of cell proportion > 1.5 when, patient prognosis bona, when phenotype is CD34+CD10+CD19+CD38+
CD20-Cell proportion and phenotype be CD34-CD10+CD19+CD38+CD20+Cell proportion and when≤1.5, patient's prognosis
It is bad；
(2) CD19 is detected+CD34+CD20+、CD10strong+CD20+CD19+、CD10+CD19+CD38-One of phenotype or
It is a variety of, there is CD19+CD34+CD20+、CD10strong+CD20+CD19+、CD10+CD19+CD38-Any one thin in phenotype
Born of the same parents are judged as abnormal residual cell, judge minimal residual and its prognosis situation according to abnormal residual cell situation.
In the present invention, subscript "+" represents the positive, indicates that corresponding antigen is expressed on cell；Subscript "-" represents feminine gender,
Indicate that corresponding antigen is not expressed on cell；Subscript " strong+ " represents strong positive, indicates corresponding antigens on cell
Expression intensity is more than normal cell.
In addition, the present invention also provides a kind of for detecting the antibody compositions of B-lineage Acute Lymphocyte Leukemia minimal residual,
The antibody compositions include following component: CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45
Antibody.
Preferably, the antibody compositions include following component: CD10-PE, CD19-APC, CD20-APC-cy7, CD34-
PerCP、CD38-V450、CD45-V500。
Further, the present invention also provides the antibody compositions in preparation leukaemia minimal residual detection reagent
Using.
The beneficial effects of the present invention are:
(1) detection kit provided by the invention includes CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38
The combination of 6 kinds of monoclonal antibodies of antibody and CD45 antibody, it is true by screening in conjunction with fluorescent marker and FCM analysis technology
The combination of fixed specific fluorescein labelled antibody, urgency can rapidly and accurately be identified by realizing one-time detection using flow cytometer
Property bone-marrow-derived lymphocyte leukaemia minimal residual abnormal cell is horizontal and its parting；Significantly improve the white blood of acute B lymphocyte
The recall rate of the minimal residual of disease, effectively reduces omission factor；
(2) in addition to the on-fixed evaluation index of single phenotype cells, the present invention also provides by using CD34+CD10+
CD19+CD38+CD20-Phenotype cells ratio and CD34-CD10+CD19+CD38+CD20+Two fixations of the sum of phenotype cells ratio refer to
Evaluation index of the mark joint as minimal residual disease, significantly improves the recall rate of patient's minimal residual disease, can be more effectively
Carry out the prognosis evaluation of patient.
The Testing index of detection kit provided by the invention can be used as intermediate result, more intuitively judge acute B
The prognosis of lymphocytic leukemia patient and situation is lapsed to, there is important directive significance for the formulation of clinical treatment,
Be conducive to improve the complete remission rate of patient.
Detailed description of the invention
The position of lymphocyte door when Fig. 1 is FCM analysis in the embodiment of the present invention 2；
When Fig. 2 is the sample of bone marrow detection in the embodiment of the present invention 3 for patient A, there is CD34+CD10+CD19+CD38+
CD20-The FCM analysis result of the cell of phenotype；Accurate circle takes CD34+(P2), CD10+(P3), CD19+(P4),
CD38+(P5), CD20-(P6), are calculated CD34 using union+CD10+CD19+CD38+CD20-Cell proportion.
When Fig. 3 is the sample of bone marrow detection in the embodiment of the present invention 3 for patient A, there is CD34-CD10+CD19+CD38+
CD20+The FCM analysis of the cell of phenotype is as a result, accurate circle takes CD34-(P2), CD10+(P3), CD19+(P4),
CD38+(P5), CD20+(P6), are calculated CD34 using union-CD10+CD19+CD38+CD20+Ratio.
When Fig. 4 is the blood sample detection in the embodiment of the present invention 3 for patient B, there is CD19+CD34+CD20+Phenotype
Abnormal residual cell FCM analysis result.
When Fig. 5 is the sample of bone marrow detection in the embodiment of the present invention 3 for patient C, there is CD10strong+CD20+CD19+
The FCM analysis result of the abnormal residual cell of phenotype.
When Fig. 6 is the sample of bone marrow detection in the embodiment of the present invention 3 for patient D, there is CD10+CD19+CD38-Phenotype
Abnormal residual cell FCM analysis result.
Specific embodiment
The preferred embodiment of the present invention is described in detail below in conjunction with embodiment.It will be appreciated that following real
Providing merely to play the purpose of explanation for example is applied, is not used to limit the scope of the present invention.The skill of this field
Art personnel without departing from the spirit and purpose of the present invention, can carry out various modifications and replace to the present invention.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified, wherein glimmering
Monoclonal antibody CD10-PE, CD19-APC, CD20-APC-cy7, CD34-PerCP, CD38-V450, CD45- of light element label
V500 is purchased from BD company；Erythrocyte cracked liquid is purchased from BD company.
The preparation of 1 B-lineage Acute Lymphocyte Leukemia minimal residual detection kit of embodiment
1, antibody compositions: the composition of antibody compositions is as follows: CD10-PE, CD19-APC, CD20-APC-cy7, CD34-
PerCP,CD38-V450,CD45-V500.I.e. by PE, APC, APC-cy7, PerCP, V450 and V500 fluorescence is successively respectively adopted
The monoclonal antibody composition of anti-CD10, CD19, CD20, CD34, CD38 and CD45 of element label.
2, BD company, including following component: ammonium chloride, potassium dihydrogen phosphate, ethylenediamine tetra-acetic acid erythrocyte cracked liquid: are purchased from
Disodium, paraformaldehyde.
3, antibody dilutes reagent: the composition that antibody dilutes reagent is as follows:
Fetal calf serum 1.5% (quality volumn concentration)；
Phosphate buffer 0.01M；
PH is 7.2~7.4.
Antibody dilution reagent the preparation method is as follows: weighing 0.1g~0.2g fetal calf serum, to be dissolved in 10ml high-temperature sterilization cold
But the 0.01M phosphate buffer after, pH are 7.2~7.4.
4, phosphate buffer (PBS): 0.01M, pH 7.2-7.4, including following component: potassium dihydrogen phosphate 1.8mmol/
L, disodium hydrogen phosphate 10mmol/L, sodium chloride 137mmol/L, potassium chloride 2.7mmol/L.
The detection method of 2 B-lineage Acute Lymphocyte Leukemia minimal residual of embodiment
It is specific as follows the present embodiment provides the detection method of detection kit prepared by above-described embodiment 1:
(1) two streaming pipe numbers are taken, one is control tube, and another pipe is detection pipe, by antibody CD10-PE, CD19-
APC, CD20-APC-cy7, CD34-PerCP, CD38-V450, CD45-V500 are added in detection pipe, guarantee CD10-PE,
The mass ratio of CD19-APC, CD20-APC-cy7, CD34-PerCP, CD38-V450, CD45-V500 are 2:5:5:13:20:40.
Antibody total amount be 16 μ l, wherein 4 μ l of CD10-PE, 2 CD19-APC μ l, 2 CD20-APC-cy7 μ l, 4 CD34-PerCP μ l,
Antibody dilution examination can be used to guarantee antibody mass ratio and volume being added in 2 μ l of CD38-V450,2 CD45-V500 μ l when necessary
Agent is to antibody beforehand dilution；Any antibody is not added in control tube.
(2) PBS diluted sample is used, so that the cell concentration of sample is 2 × 106/ mL, step (1) detection pipe and
Sample after being separately added into 500 μ l dilution in control tube；
(3) low speed mixes, and room temperature is protected from light incubation 15 minutes；
(4) it is separately added into 1ml erythrocyte cracked liquid into control tube and detection pipe, mixes, room temperature is protected from light 8 minutes；
(5) it low speed centrifuge 300rct/ minutes, is centrifuged 5 minutes, removes supernatant；
(6) it is separately added into 1ml PBS in the control tube and detection pipe in above-mentioned steps (5), after mixing, 150rct/ points
Clock is centrifuged 5 minutes, removes supernatant；
(7) it is to be measured that 150 μ l PBS resuspension is separately added into the control tube and detection pipe of step (6)；
(8) machine testing on is located at lymphocyte populations as shown in Figure 1 according to the instrument condition that each quality-control product regulates
The position P1, and 1,000,000 signals are obtained, it is analyzed using BD DivaSoftware.
(9) data analysis and result judgement: one of following index or a variety of is detected, is judged according to testing result small
Residual Disease and its prognosis situation: (1) phenotype is CD34+CD10+CD19+CD38+CD20-Cell ratio and phenotype be
CD34-CD10+CD19+CD38+CD20+Cell ratio, calculate sum of the two；It (2) is CD19 with the presence or absence of phenotype+CD34+
CD20+Abnormal residual cell；It (3) is CD10 with the presence or absence of phenotypestrong+CD20+CD19+Abnormal residual cell；(4) whether
It is CD10 there are phenotype+CD19+CD38-Abnormal residual cell.
3 B-lineage Acute Lymphocyte Leukemia minimal residual detection kit of embodiment and its detection method are examined in clinical sample
Application in survey
Using the detection kit of embodiment 1 and the detection method of embodiment 2 respectively to 127 parts from different acute Bs
The clinical samples such as marrow, the blood of lymphocytic leukemia patient detect, and the clinical sample detection of each patient is respectively provided with
Test group and control group, test group is using the kit of embodiment 1 and the detection method of embodiment 2, and control group is using control examination
Agent box and control test method are detected.
Contrast agents box is only that antibody compositions are different from the difference of the kit of embodiment 1, contrast agents box antibody
Composition component is as follows: CD10-PE, CD19-APC, CD34-PerCP, CD38-V450, CD45-V500；Control test method with
The difference of the detection method of embodiment 2 is only that the antibody compositions being added in step (1) are different: 4 μ l of CD10-PE, CD19-
2 μ l of APC, 4 CD34-PerCP μ l, 2 CD38-V450 μ l, 2 CD45-V500 μ l, the antibody total amount of addition are 14 μ l, are guaranteed
CD10 antibody, CD19 antibody, CD34 antibody, CD38 antibody and CD45 antibody mass ratio be 2:5:13:20:40.
Clinical sample testing result shows that test group can accurately obtain CD34+CD10+CD19+CD38+CD20-Cell
Ratio and phenotype are CD34-CD10+CD19+CD38+CD20+The sum of cell proportion；And phenotype is CD19+CD34+CD20+
Abnormal residual cell, phenotype CD10strong+CD20+CD19+Abnormal residual cell；Phenotype is CD10+CD19+CD38-'s
The information of abnormal residual cell, and then there is higher minimal residual recall rate, it can more precisely judge minimal residual disease
Situation and its prognosis situation；And control group kit then can not accurately obtain CD34+CD10+CD19+CD38+CD20-Cell ratio
Example and CD34-CD10+CD19+CD38+CD20+Cell proportion, it is pre- for the recall rate and disease of minimal residual abnormal cell
The accuracy predicted afterwards is significantly lower than test group.
The FCM analysis result of partial clinical sample as figures 2-6, the wherein CD34 of the bone marrow specimens of patient A+
CD10+CD19+CD38+CD20-Cell proportion Indexs measure result as shown in Fig. 2, CD34-CD10+CD19+CD38+CD20+'s
Cell proportion Indexs measure result through software as shown in figure 3, calculate, the CD34 of patient A+CD10+CD19+CD38+CD20-Cell
Ratio and CD34-CD10+CD19+CD38+CD20+The sum of cell proportion be 5.22, judge its prognosis bona according to testing result；
The CD19 of the blood sample of patient B+CD34+CD20+Abnormal residual cell Indexs measure result is as shown in Figure 4, the results showed that, suffer from
Person B exists abnormal；The CD10 of the bone marrow specimens of patient Cstrong+CD20+CD19+The testing result of abnormal residual cell index is as schemed
Shown in 5, the results showed that, patient C exists abnormal；The CD10 of the bone marrow specimens of patient D+CD19+CD38-Abnormal residual cell index
Testing result is as shown in Figure 6, the results showed that, patient D exists abnormal.
Although above the present invention is described in detail with a general description of the specific embodiments, at this
On the basis of invention, it can be modified or is improved, this will be apparent to those skilled in the art.Therefore,
These modifications or improvements without departing from theon the basis of the spirit of the present invention are fallen within the scope of the claimed invention.
Claims (10)
1. a kind of detection kit of B-lineage Acute Lymphocyte Leukemia minimal residual, which is characterized in that the kit includes
CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody.
2. kit according to claim 1, which is characterized in that the CD10 antibody, CD19 antibody, CD20 antibody,
CD34 antibody, CD38 antibody and CD45 antibody contain different fluorescein labels；
Preferably, the fluorescein of the CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody
Label is respectively any one in PE, APC, APC-cy7, PerCP, V450, V500.
3. kit according to claim 1 or 2, which is characterized in that the CD10 antibody, CD19 antibody, CD20 antibody,
CD34 antibody, CD38 antibody and CD45 antibody by from front to back sequence successively by PE, APC, APC-cy7, PerCP, V450,
V500 fluorescein label.
4. described in any item kits according to claim 1~3, which is characterized in that the CD10 antibody, CD19 antibody,
The antibody mass ratio of CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody when detecting for minimal residual is as follows:
The mass ratio of CD10 antibody, CD20 antibody and CD38 antibody is 2:5:20~1:5:5；
And/or
The mass ratio of CD34 antibody and CD19 antibody is 3:1~1:2；
And/or
The mass ratio of CD45 antibody and CD19 antibody is 4:1~8:1；
Preferably, the mass ratio of CD10 antibody, CD20 antibody and CD38 antibody is 2:5:20~1:5:8；
And/or
The mass ratio of CD34 antibody and CD19 antibody is 3:1~2:1；
And/or
The mass ratio of CD45 antibody and CD19 antibody is 6:1~8:1.
5. kit according to any one of claims 1 to 4, which is characterized in that the CD10 antibody, CD19 antibody,
CD20 antibody, CD34 antibody, the mass ratio of CD38 antibody and CD45 antibody when detecting for minimal residual are 2:5:5:13:
20:40。
6. described in any item kits according to claim 1~5, which is characterized in that the kit is being used for minimal residual
To be detected using stream type cell analyzer when detection, according to the obtained fluorescein-labeled fluorescence signal intensity, really
Determine the cell proportion that one of CD10, CD19, CD20, CD34, CD38, CD45 or a variety of are expressed as positive or negative, judges
Minimal residual situation.
7. described in any item kits according to claim 1~6, which is characterized in that the kit is being used for minimal residual
Working procedure when detection includes the following steps:
(1) sample is acquired, sample is pre-processed；
(2) be added in the sample the fluorescein-labeled CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody,
CD38 antibody and CD45 antibody carry out the fluorescence antibody label of sample；
(3) after carrying out erythrocyte splitting processing to the sample, using flow cytomery；
(4) according to flow cytomery as a result, determining one of CD10, CD19, CD20, CD34, CD38, CD45 or a variety of
It is expressed as the cell proportion of positive or negative, judges minimal residual situation；
Preferably, the sample includes being selected from one of marrow, blood.
8. kit according to claim 6 or 7, which is characterized in that the judgment method of the minimal residual situation is choosing
The one or more of method are descended freely:
(1) detection phenotype is CD34 respectively+CD10+CD19+CD38+CD20-Cell proportion and phenotype be CD34-CD10+CD19+
CD38+CD20+Cell proportion, according to phenotype be CD34+CD10+CD19+CD38+CD20-Cell proportion and phenotype be CD34-
CD10+CD19+CD38+CD20+The outer minimal residual of the sum of cell proportion judgement and its prognosis situation；
Preferably, when phenotype is CD34+CD10+CD19+CD38+CD20-Cell proportion and phenotype be CD34-CD10+CD19+
CD38+CD20+The sum of cell proportion > 1.5 when, patient prognosis bona, when phenotype is CD34+CD10+CD19+CD38+CD20-
Cell proportion and phenotype be CD34-CD10+CD19+CD38+CD20+Cell proportion and when≤1.5, patient's prognosis mala；
(2) CD19 is detected+CD34+CD20+、CD10strong+CD20+CD19+、CD10+CD19+CD38-One of phenotype is more
Kind, there is CD19+CD34+CD20+、CD10strong+CD20+CD19+、CD10+CD19+CD38-The cell of any one in phenotype
It is judged as abnormal residual cell, minimal residual and its prognosis situation is judged according to abnormal residual cell situation.
9. a kind of for detecting the antibody compositions of B-lineage Acute Lymphocyte Leukemia minimal residual, which is characterized in that the combination
Object includes following component: CD10 antibody, CD19 antibody, CD20 antibody, CD34 antibody, CD38 antibody and CD45 antibody；
Preferably, the antibody compositions include following component: CD10-PE, CD19-APC, CD20-APC-cy7, CD34-
PerCP、CD38-V450、CD45-V500。
10. application of the antibody compositions as claimed in claim 9 in preparation leukaemia minimal residual detection reagent.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN112578117A (en) * | 2021-02-22 | 2021-03-30 | 信纳克(北京)生化标志物检测医学研究有限责任公司 | Antibody composition and application thereof in screening post-transplantation lymphocyte proliferative diseases |
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| CN111912983A (en) * | 2020-06-24 | 2020-11-10 | 北京积水潭医院 | Antibody composition for detecting minimal residual disease of multiple myeloma, kit and application |
| CN112578117A (en) * | 2021-02-22 | 2021-03-30 | 信纳克(北京)生化标志物检测医学研究有限责任公司 | Antibody composition and application thereof in screening post-transplantation lymphocyte proliferative diseases |
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| CN112552407A (en) * | 2021-02-24 | 2021-03-26 | 信纳克(北京)生化标志物检测医学研究有限责任公司 | Antibody composition and application thereof in detecting acute B lymphocyte leukemia |
| CN112552407B (en) * | 2021-02-24 | 2021-05-25 | 信纳克(北京)生化标志物检测医学研究有限责任公司 | Antibody composition and application thereof in detecting acute B lymphocyte leukemia |
| CN113419066A (en) * | 2021-08-23 | 2021-09-21 | 信纳克(北京)生化标志物检测医学研究有限责任公司 | Reagent composition for screening and/or diagnosing clonal diseases by one-step method and application thereof |
| CN113419066B (en) * | 2021-08-23 | 2021-11-02 | 信纳克(北京)生化标志物检测医学研究有限责任公司 | Reagent composition for screening and/or diagnosing clonal diseases by one-step method and application thereof |
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| CN113933511B (en) * | 2021-09-18 | 2024-03-22 | 广州金域医学检验中心有限公司 | Antibody composition and method for detecting acute B lymphocyte leukemia tiny residue |
| CN117169517A (en) * | 2023-11-03 | 2023-12-05 | 赛德特(北京)生物工程有限公司 | Method and kit for detecting CD28 antibody residues in T lymphocyte preparation |
| CN117169517B (en) * | 2023-11-03 | 2024-01-19 | 赛德特(北京)生物工程有限公司 | Method and kit for detecting CD28 antibody residues in T lymphocyte preparation |
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