CN206529458U - A kind of kit detected for T cell Minimal Residual Disease of Leukemia - Google Patents
A kind of kit detected for T cell Minimal Residual Disease of Leukemia Download PDFInfo
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- CN206529458U CN206529458U CN201720025366.3U CN201720025366U CN206529458U CN 206529458 U CN206529458 U CN 206529458U CN 201720025366 U CN201720025366 U CN 201720025366U CN 206529458 U CN206529458 U CN 206529458U
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- reagent bottle
- box body
- minimal residual
- standing groove
- residual disease
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Abstract
The utility model discloses a kind of kit detected for T cell Minimal Residual Disease of Leukemia, including box body, lid is hinged with the box body, the upper center of the lid is provided with stopper slot, handle is rotatably connected in the stopper slot, the both sides of the box body are equipped with opening, run through in the opening provided with placement plate, the both sides for placing plate are equipped with sliding block, relative side wall is provided with chute corresponding with sliding block in the box body, and sliding block is located in chute, the placement plate is provided with standing groove, multiple suckers are equidistantly provided with the standing groove, the sucker is provided with gas vent, the gas vent is provided with sealing-plug, the upper end of the sucker is fixed with support bar.The utility model realizes the gripping to different reagent bottles by the combination of fixing pipe, screw rod and sucker, while different types of reagent bottle can also be stored, convenient operation is also convenient for carrying, suitable to promote.
Description
Technical field
The utility model is related to biological technical field, more particularly to a kind of T cell Minimal Residual Disease of Leukemia that is used for is detected
Kit.
Background technology
ALL (ALL) is one of most common adult acute leukemia, and it is that a class is drenched with former children
Malignant hematologic disease with the characteristics of bar undesired cell proliferation, accumulation, infiltration.Multinomial research report uses systematic treating scheme, and ALL suffers from
Person's complete remission rate (CR) 70%-90%, 3-5 disease-free survival rate is 60%.Although the first of adult ALL controls CR rates oneself has bright
Aobvious raising, but high recurrence rate, long-term survival rate are still relatively low.Equally, children acute myelocytic leukemia accounts for acute leukemia
20% or so, but account for children AL and die of illness more than the 50% of number.At present using newtype drug and the elder generation of HSCT
Entering therapeutic scheme can make patient obtain the long-term disease-free survival close to 70% and 5 years Event-free survival rates more than 50%, still
Recurrence rate and case fatality rate are equally higher.The main cause for occurring similar above Recurrent death is exactly that minimal residual (MRD) is drawn
Rise.ALL is in morbidity about 10 in body12The leukaemia of left and right, but pass through Treated with Chemotherapeutic Drugs thing
Use in conjunction, when up to complete incidence graph, in vivo still residue 108-1010The leukaemia of left and right, but these remaining leukaemia
By existing morphologic detection, oneself can not make accurate test and appraisal to cell, and these remaining leukaemias are leukemia relapse
Root, therefore the state of still remaining leukaemia is referred to as microresidual disease in vivo after this leukaemia complete incidence graph.
Current MRD detection techniques are mainly flow cytometry and polymerase chain reaction technology.Although PCR sensitivity is high,
But it is less available for the fusion that PCR is monitored, use narrow range；Although it is high that gene mutation, proto-oncogene are overexpressed coverage rate,
But sensitivity is low.Although mpFC is 10 for the detection sensitivity of recurrent disease-4, but complicated multidimensional data is depended on
Experimenter analyzes, and human factor influence is big, is unfavorable for clinical criteriaization detection.In addition, after chemotherapy leukemia antigen table
There is interference effect to MRD mpFC detections up to level.Detection MRD sensitivity can be improved dependent on Molecular tools, can be with
Reach 10-5；The retracing sequence that diversity is enriched is amplified however, real-time quantitative PCR need to design special primer according to patient
Come, its testing cost costliness, labour intensive, be hardly formed standardized assay flow, at present when being detected more use and try
Agent box is detected, but existing detection box, simple in construction, can only play carrying, it is impossible to according to the size of reagent bottle
It is adjusted, and reagent bottle can not be stored, it is very inconvenient, therefore, being used for T cell leukaemia minimal residual we have proposed one kind
The kit of disease detection solves the above problems.
Utility model content
The purpose of this utility model is that, in order to solve shortcoming present in prior art, and the one kind proposed is used for T cell
The kit of Minimal Residual Disease of Leukemia detection.
To achieve these goals, the utility model employs following technical scheme：
Box is hinged with a kind of kit detected for T cell Minimal Residual Disease of Leukemia, including box body, the box body
Lid, the upper center of the lid, which is provided with stopper slot, the stopper slot, is rotatably connected to handle, and the both sides of the box body are all provided with
Have and run through in opening, the opening provided with plate is placed, the both sides of the placement plate are equipped with side relative in sliding block, the box body
Wall is provided with chute corresponding with sliding block, and sliding block is located in chute, and the placement plate is provided with standing groove, the standing groove
Multiple suckers are equidistantly provided with, the sucker is provided with gas vent, and the gas vent is provided with sealing-plug, the upper end of the sucker
Support bar is fixed with, the one end of the support bar away from sucker is fixed with loading plate, the loading plate and is fixed with reagent bottle clip
Tight device, the bottom that the side of the placement plate is provided with handle, the box body is equidistantly provided with two dividing plates, two from left to right
Individual dividing plate by the equal-sized first reagent bottle storage area of inside equally-spaced three of box body, the second reagent bottle storage area and
3rd reagent bottle storage area, the bottom of three reagent bottle storage areas is equidistantly provided with multiple reagent bottle standing grooves.
Preferably, the reagent bottle clamping device includes the fixing pipe being arranged on loading plate, and the fixing pipe is one end
The staving of closing, one end of the fixed duct occlusion is welded on loading plate, and the fixing pipe is provided with tapped through hole, the spiral shell
Run through in line through hole and be provided with screw rod, one end of the screw rod is provided with moving block, and the one end of the screw rod away from moving block, which is provided with, to be squeezed
Foam-rubber cushion is coated with briquetting, and crushing block.
Preferably, anti-slip veins are equipped with the handle and handle.
Preferably, the lid is provided with lock, and the box body is provided with and latched corresponding lockhole.
Preferably, it is equipped with heat-insulation layer on the side wall in the box body.
Preferably, the reagent bottle standing groove of the first reagent bottle storage area is more than the reagent bottle of the second reagent bottle storage area
Standing groove, the reagent bottle standing groove of the second reagent bottle storage area is more than the reagent bottle standing groove of the 3rd reagent bottle storage area.
In the utility model, in use, being opened lid using lock and lockhole, sucker is taken out, as needed from examination
Corresponding reagent bottle is taken out in agent bottle rest area, different reagent bottles can be fixedly clamped by screw rod and fixing pipe, so that
Detected, the setting of handle, which facilitates, to be carried, the utility model is realized by the combination of fixing pipe, screw rod and sucker
Gripping to different reagent bottles, while different types of reagent bottle can also store, convenient operation is also convenient for carrying, suitably
Promote.
Brief description of the drawings
Fig. 1 be the utility model proposes it is a kind of for T cell Minimal Residual Disease of Leukemia detect kit structure
Schematic diagram；
Fig. 2 be the utility model proposes it is a kind of for T cell Minimal Residual Disease of Leukemia detect kit inside
Structural representation.
In figure：1 handle, 2 stopper slots, 3 lids, 4 box bodys, 5 handles, 6 chutes, 7 dividing plates, 8 fixing pipes, 9 suckers, 10 are put
Put plate, 11 screw rods, 12 support bars, 13 sliding blocks.
Embodiment
Below in conjunction with the accompanying drawing in the utility model embodiment, the technical scheme in the utility model embodiment is carried out
Clearly and completely describe, it is clear that described embodiment is only a part of embodiment of the utility model, rather than whole
Embodiment.
A kind of reference picture 1-2, kit detected available for T cell Minimal Residual Disease of Leukemia is as follows using step：
1st, One step RT-PCR step；
2nd, magnetic beads for purifying step
1) amount for taking out magnetic bead vibration mixing 5min 45 μ l/ samples of taking-up is put in room temperature 10min；
2) RT-PCR product respectively takes out 20 μ l, remaining 5 μ ljustincase, then each sample adds 30 μ lH2O, finally
Magnetic bead is added according to 45 μ l/, is inhaled with rifle and blows about 10 mixings, be put in room temperature 2min；
3) uncap and be put in 1min or so on magnetic frame, liquid, which becomes clarification, can wash out liquid, add 125 μ l85% second
Wash out and discard after alcohol, 1min, magnetic bead is put in room temperature 8min (5-10min) and dried.
3rd, DNA concentration is determined with Nanodrop.If concentration is 20ng/ μ l, 5 times are diluted；If close to 40ng/ μ l,
10 times of dilution.
4th, end is repaired：Following component (50 μ l) is mixed in 1.5ml centrifuge tubes
1) the patient's sample genomic DNA (or moisturizing to 34 μ l) of 34 μ l purifying
2) buffer solution is repaired in 5 μ l10 × end
3) 5 μ l2.5mMdNTP mixtures
4)5μl10mMATP
5) 1 μ l ends repair enzyme
Room temperature is placed 45 minutes.Purified with QIAquickPCR purification columns, eluted with 34 μ l buffer solutions.
5th, 3 ' ends add A：Following component (50 μ l) is mixed in 1.5ml centrifuge tubes：
1) DNA after the end of 34 μ l steps 4 is repaired
2) 5 μ lKlenow buffer solutions=NEBbuffer2
3)10μl1mMdATP
4) 1 μ lKlenow fragments (5 prime excision enzyme activities of 3 ' à 5 ')
(1mMdATP with 100mMdATP mother liquors dilute, often the μ l of pipe 25 dispense, can only freeze thawing once)
Purified with MinElutePCR purification columns, the elution of 12 μ l buffer solutions.
6th, joint is connected：Following component (30ul) is mixed in 1.5ml centrifuge tubes
1) DNA of 11 μ l steps 5
2) 15 μ lDNA connection buffer solutions
3)1μl1:The linker oligonucleotides of 20 dilutions
4) 3 μ lDNA ligases
If doing multiple repetitions simultaneously, it is ensured that each parallel reaction adds different joints, marks clear.
7th, gel screening removes unnecessary joint：
The 6 μ l sample-loading buffers for diluting 10 times are added in the reaction solution of the steps of 30 μ l the 6th, loading to two E-gel glue
Kong Zhong.The 50bpDNAladder loadings for taking 20 μ l to dilute 10 times in addition.Electrophoresis 20 minutes.
DNA of the size between 150bp to 450bp is cut, is purified with QIAquick gel purification kits,
30 μ l elution buffers are eluted.
8th, Illumina primer PCRs are used：
IlluminaPCR primers 1.1 and 2.1 are carried out 1 with Gibco water：1 dilution
Following component (60ul) is mixed in PCR pipe：
1) DNA of 30 μ l steps 4
2) 28 μ lPhusion PCR premixed liquids
3) primer 1.1 of 1 μ l dilutions
4) the primer 2 .1 of 1 μ l dilutions
PCR cycle:
9th, molecular weight screening is carried out on 2% Ago-Gel：
The 1 μ l sample-loading buffers diluted are added in the PCR reaction solutions of previous step, loading is into three E-gel glue holes.
50bp the and 100bpDNAladder loadings for taking 20 μ l to dilute 10 times respectively in addition.Electrophoresis 30 minutes.Size is cut in 400bp
The DNA of left and right.Purify DNA with QIAquick gel purification kits, eluted with 30 μ l elution buffers.
10th, DNA concentration is determined.Gained DNA concentration is determined with NanoDrop.
In the utility model, in use, using lock and lockhole lid 3 is opened, sucker 9 is taken out, as needed from
Corresponding reagent bottle is taken out in reagent bottle rest area, can be solid to be clamped to different reagent bottles by screw rod 11 and fixing pipe 8
Fixed, so as to be detected, the setting of handle 1, which facilitates, to be carried.
It is described above, only the utility model preferably embodiment, but protection domain of the present utility model is not
This is confined to, any one skilled in the art is in the technical scope that the utility model is disclosed, according to this practicality
New technical scheme and its utility model design are subject to equivalent substitution or change, should all cover in protection model of the present utility model
Within enclosing.
Claims (6)
1. a kind of kit detected for T cell Minimal Residual Disease of Leukemia, including box body (4), it is characterised in that the box
Lid (3) is hinged with body (4), the upper center of the lid (3) is provided with the company of rotation in stopper slot (2), the stopper slot (2)
Handle (1) is connected to, the both sides of the box body (4), which are equipped with opening, the opening, to be run through provided with plate (10) is placed, described to place
The both sides of plate (10) are equipped with side wall relative in sliding block (13), the box body (4) and are provided with and the corresponding chute of sliding block (13)
(6), and sliding block (13) is located in chute (6), the placement plate (10) is provided with standing groove, is equidistantly provided with the standing groove
Multiple suckers (9), the sucker (9) is provided with gas vent, and the gas vent is provided with sealing-plug, the upper end of the sucker (9)
Support bar (12) is fixed with, the one end of the support bar (12) away from sucker (9) is fixed with loading plate, the loading plate fixed
There is a reagent bottle clamping device, the side of the placement plate (10) is provided with bottom in handle (5), the box body (4) from left to right
Equidistantly provided with two dividing plates (7), two dividing plates (7) are by the inside equally-spaced three equal-sized first of box body (4)
Reagent bottle storage area, the second reagent bottle storage area and the 3rd reagent bottle storage area, bottom of three reagent bottle storage areas etc.
Spacing is provided with multiple reagent bottle standing grooves.
2. a kind of kit detected for T cell Minimal Residual Disease of Leukemia according to claim 1, its feature exists
In the reagent bottle clamping device includes the fixing pipe (8) being arranged on loading plate, and the fixing pipe (8) is what one end was closed
Staving, one end of fixing pipe (8) closing is welded on loading plate, and the fixing pipe (8) is provided with tapped through hole, the spiral shell
Run through in line through hole and be provided with screw rod (11), one end of the screw rod (11) is provided with moving block, and the screw rod (11) is away from moving block
One end provided with being coated with foam-rubber cushion on crushing block, and crushing block.
3. a kind of kit detected for T cell Minimal Residual Disease of Leukemia according to claim 1, its feature exists
In being equipped with anti-slip veins on the handle (1) and handle (5).
4. a kind of kit detected for T cell Minimal Residual Disease of Leukemia according to claim 1, its feature exists
In the lid (3) is provided with and latched corresponding lockhole provided with lock, the box body (4).
5. a kind of kit detected for T cell Minimal Residual Disease of Leukemia according to claim 1, its feature exists
In being equipped with heat-insulation layer on the side wall in the box body (4).
6. a kind of kit detected for T cell Minimal Residual Disease of Leukemia according to claim 1, its feature exists
In the reagent bottle standing groove of the first reagent bottle storage area is more than the reagent bottle standing groove of the second reagent bottle storage area, described
The reagent bottle standing groove of second reagent bottle storage area is more than the reagent bottle standing groove of the 3rd reagent bottle storage area.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720025366.3U CN206529458U (en) | 2017-01-10 | 2017-01-10 | A kind of kit detected for T cell Minimal Residual Disease of Leukemia |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720025366.3U CN206529458U (en) | 2017-01-10 | 2017-01-10 | A kind of kit detected for T cell Minimal Residual Disease of Leukemia |
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| Publication Number | Publication Date |
|---|---|
| CN206529458U true CN206529458U (en) | 2017-09-29 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201720025366.3U Active CN206529458U (en) | 2017-01-10 | 2017-01-10 | A kind of kit detected for T cell Minimal Residual Disease of Leukemia |
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| Country | Link |
|---|---|
| CN (1) | CN206529458U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114620330A (en) * | 2022-03-14 | 2022-06-14 | 山东省物化探勘查院 | Data real-time recording device for geographic mapping |
-
2017
- 2017-01-10 CN CN201720025366.3U patent/CN206529458U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114620330A (en) * | 2022-03-14 | 2022-06-14 | 山东省物化探勘查院 | Data real-time recording device for geographic mapping |
| CN114620330B (en) * | 2022-03-14 | 2023-09-05 | 山东省物化探勘查院 | Data real-time recording device for geographical mapping |
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