CN215362482U - TCR-T cell detection kit - Google Patents

Abstract

The utility model belongs to the technical field of peripheral blood TCR-T cell detection equipment, and discloses a TCR-T cell detection kit, which aims to solve the problems of more reagents and inconvenience in carrying and storage in a flow detection method in the prior art. The utility model has simple integral structure and convenient carrying, thereby being convenient for detecting the TCR-T cells of the peripheral blood.

Description

TCR-T cell detection kit

Technical Field

The utility model belongs to the technical field of peripheral blood TCR-T cell detection equipment, and particularly relates to a TCR-T cell detection kit.

Background

TCR-T immunotherapy is characterized by that the intracellular signal region containing CD3 zeta of TCR capable of recognizing a certain tumor antigen and the costimulatory molecule are coupled into a chimeric protein, and the T cell of patient is transfected by means of gene transduction, so that it can express chimeric antigen receptor (TCR). After the T cells of the patient are recoded, a large number of TCR-T cells with tumor specificity are generated, and the high-quality TCR-T cells which are expanded in vitro are returned to the body of the patient to kill cancer cells in a targeted manner. The survival and proliferation of TCR-T cells after reinfusion into a patient are closely related to the therapeutic effect and related side effects, so that TCR-T cell monitoring of the blood of the patient after reinfusion is an indispensable project.

The prior follow-up detection of TCR-T cells in peripheral blood can be realized by a flow-type and fluorescence quantitative PCR method. The fluorescent quantitative PCR method has the disadvantages of complex operation, slow detection speed, low efficiency, multiple reagents and incapability of obtaining the absolute content of TCR-T cells. The flow detection method can be used for quickly, efficiently, simply and conveniently detecting TCR-T cells in peripheral blood more accurately. In the flow detection method, a plurality of reagents such as erythrocyte lysis, buffer solution, flow antibodies and the like are involved, the preparation is complicated, and the carrying is inconvenient.

SUMMERY OF THE UTILITY MODEL

In order to solve the problems of more reagents and inconvenient carrying and storage in the prior art by adopting a flow detection method, the utility model provides a TCR-T cell detection kit which is used for detecting TCR-T cells in peripheral blood of a clinical patient.

The technical scheme adopted by the utility model is as follows: the utility model provides a TCR-T cell detect reagent box, its includes the kit body, this internal first storage tank that is used for placing buffer solution reagent bottle, the second storage tank that is used for placing lysate reagent bottle and the third storage tank that is used for placing antibody reagent bottle that is equipped with of kit, and kit body top still articulates and is equipped with the apron, and the apron passes through the buckle with the kit body and is connected the realization closure.

Optionally, a cold insulation bag placing cavity and a sealing cover used for sealing the cold insulation bag placing cavity are formed in one side of the kit body, and the cold insulation bag placing cavity is communicated with the first storage groove, the second storage groove and the third storage groove respectively.

Optionally, a connecting channel is further sequentially communicated among the first storage groove, the second storage groove and the third storage groove.

Optionally, one end of the sealing cover is hinged to the side wall of the kit body, and the other end of the sealing cover is connected through a buckle to seal the cold-insulation bag placing cavity.

Optionally, the top of the kit body is further provided with a step wall, the first storage tank, the second storage tank and the third storage tank are all arranged below the step wall, and when the cover plate is closed, the upper part of the step wall is attached to the cover plate; the height of the step wall is matched with the thicknesses of bottle caps of a buffer solution reagent bottle, a lysate reagent bottle and an antibody reagent bottle.

Optionally, the bottoms of the first storage tank, the second storage tank and the third storage tank are provided with cushion pads.

Optionally, the first storage slot, the second storage slot and the third storage slot are all obliquely arranged.

Optionally, a clamping end is arranged on one side of the cover plate, a first handle is arranged on the side face of the kit body, and a buckling end matched with the clamping end is arranged at the top of the first handle.

Optionally, a second handle is further arranged on the reagent box body and located on the opposite side of the first handle.

Optionally, the buffer solution reagent bottle, the lysate reagent bottle and the antibody reagent bottle are all brown reagent bottles, and labels corresponding to the buffer solution reagent bottle, the lysate reagent bottle and the antibody reagent bottle are pasted outside the brown reagent bottles.

The utility model has the beneficial effects that:

according to the utility model, the buffer solution reagent bottle, the lysate reagent bottle and the antibody reagent bottle are stored in the TCR-T cell detection kit, and the temperature of the inner cavity of the kit can be regulated to 2-8 ℃, so that the reagents can be conveniently stored. And each reagent in the kit can be taken quickly and conveniently, and the whole kit has the advantages of convenience in carrying and simplicity in operation, so that the detection of TCR-T cells in peripheral blood is facilitated.

The advantages of the utility model are not limited to this description, but are described in more detail in the detailed description for better understanding.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.

FIG. 1 is a schematic structural diagram of the present invention in one embodiment;

FIG. 2 is a schematic cross-sectional view taken at A-A in FIG. 1;

FIG. 3 is a schematic structural diagram of the present invention in another embodiment;

in the figure: the kit comprises a kit body, a cover plate 2, a first storage groove 3, a second storage groove 4, a third storage groove 5, a cold insulation bag placing cavity 6, a sealing cover 7, a step wall 11, a buckling end 12, a clamping end 21 and a connecting channel 100.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.

Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the utility model, as claimed, but is merely representative of selected embodiments of the utility model. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

In the description of the embodiments of the present invention, it should be noted that the indication of the orientation or the positional relationship is based on the orientation or the positional relationship shown in the drawings, or the orientation or the positional relationship which is usually placed when the utility model is used, or the orientation or the positional relationship which is conventionally understood by those skilled in the art, is only for the convenience of describing the present invention and simplifying the description, and does not indicate or imply that the indicated device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first" and "second" are used merely to distinguish one description from another, and are not to be construed as indicating or implying relative importance.

In the description of the embodiments of the present invention, it should be further noted that the terms "disposed" and "connected" are to be interpreted broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected, unless explicitly stated or limited otherwise; may be directly connected or indirectly connected through an intermediate. For those skilled in the art, the technical solutions in the embodiments of the present invention will be clearly and completely described by understanding the specific meanings of the above terms in the present invention according to the specific situations and the drawings of the specification, and it is obvious that the described embodiments are a part of the embodiments of the present invention, and not all of the embodiments. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.

The utility model is further illustrated with reference to the accompanying drawings and specific examples.

As shown in fig. 1-2, the utility model provides a TCR-T cell detection kit, which comprises a cuboid kit body 1, wherein a first storage tank 3 for storing buffer solution reagent bottles, a second storage tank 4 for storing lysate reagent bottles, and a third storage tank 5 for storing antibody reagent bottles are arranged in the kit body 1, the top of the kit body 1 is further hinged with a cover plate 2, and the cover plate 2 is connected with the kit body 1 through a buckle to realize closing.

The buffer solution reagent bottle contains a buffer solution for washing the antibody and the cell debris, and the buffer solution reagent bottle is used for stopping cell lysis, washing away redundant antibody and cell debris, and resuspending the stained cells for subsequent flow detection. The lysate reagent bottle contains erythrocyte lysate, and the function of the lysate reagent bottle is to lyse erythrocytes in a sample after the antibody incubation, and the residual leukocytes are used for flow detection. The antibody reagent bottle contains corresponding marker antibodies with dyes with different wavelengths, namely anti-humanCD 3-APC and anti-TCR-PE, which are used for marking TCR-T cells with corresponding antibodies, and then detecting the TCR-T cells in peripheral blood by a flow method.

The prepared buffer solution reagent bottle, lysate reagent bottle and antibody reagent bottle are respectively put into a first storage tank 3, a second storage tank 4 and a third storage tank 5 corresponding to the kit body 1, in the specific TCR-T cell detection process, a certain volume of blood sample to be detected is taken by a pipettor and added into an EP tube, the antibody in the antibody reagent bottle is used for marking the sample to be detected, the lysate in the lysate reagent bottle is used for cracking the blood sample, the buffer solution in the buffer solution reagent bottle is used for stopping cracking, the buffer solution in the buffer solution reagent bottle is used for centrifuging after standing, supernatant is removed, the buffer solution in the buffer solution reagent bottle is used for cleaning cell sediment, after centrifuging, the buffer solution in the buffer solution reagent bottle is used for re-suspending the cell sediment again, and the flow type instrument is used for detection. The kit greatly facilitates the storage and the taking of buffer solution, lysate and antibody, and has the advantages of convenient carrying, safe storage, simple and convenient operation and the like.

Because the transportation and storage of the buffer solution, the lysate and the antibody need to be controlled at the temperature of 2-8 ℃, in order to reduce the temperature in the kit body 1, in another embodiment, a cold-insulation bag placing cavity 6 and a sealing cover 7 for sealing the cold-insulation bag placing cavity 6 are formed in one side of the kit body 1, and the cold-insulation bag placing cavity 6 is respectively communicated with the first storage tank 3, the second storage tank 4 and the third storage tank 5 through connecting channels 100. The cold insulation bag or other independent temperature control objects are placed in the cold insulation bag placing cavity 6, and due to heat transfer, the temperature of the inner cavity of the kit body 1 can be reduced to 2-8 ℃. In order to better realize temperature control in the inner cavity of the kit body 1, a temperature sensor is further arranged in the inner cavity of the kit body 1, the temperature of the inner cavity of the kit body 1 can be monitored through the temperature sensor, a controller, a display and an alarm are further arranged outside the kit body 1, the temperature sensor, the display and the alarm are respectively and electrically connected with the controller, and the alarm gives an alarm to remind of controlling the temperature of the inner cavity of the kit body 1 after the temperature monitored by the temperature sensor exceeds the range of 2-8 ℃; the display can display the temperature of the inner cavity of the kit body 1 in real time, so that the regulation and control of the temperature of the inner cavity of the kit body 1 are conveniently controlled.

In another embodiment, the first storage compartment 3, the second storage compartment 4 and the third storage compartment 5 are sequentially communicated with a connecting passage 100, so as to further enhance the temperature balance among the first storage compartment 3, the second storage compartment 4 and the third storage compartment 5. Because buffer solution reagent bottle, lysate reagent bottle and antibody reagent bottle put into first holding tank 3, second holding tank 4 and third holding tank 5 back respectively, the reagent bottle lateral wall can be hugged closely with the holding tank, can carry out the shutoff with connecting channel 100, consequently, need be in advance with 1 inner chamber temperature regulation of kit body suitable before putting into buffer solution reagent bottle, lysate reagent bottle and antibody reagent bottle.

In order to adjust the peripheral temperature of the reagent bottles when buffer solution reagent bottles, lysate reagent bottles and antibody reagent bottles are put in, in another embodiment, as shown in fig. 3, annular channels are arranged in the middle of the first storage tank 3, the second storage tank 4 and the third storage tank 5, and the annular channels are communicated with the cold-insulation bag placing cavity 6, so that the temperature of the inner cavity of the reagent kit body 1 can be better adjusted. More preferably, the annular passages in the middle of the first storage tank 3, the second storage tank 4 and the third storage tank 5 are all communicated with each other.

In order to further guarantee the temperature of the inner cavity of the kit body 1, the kit body 1 is also externally provided with a packing box for better sealing the kit body 1.

In order to facilitate the opening and closing of the sealing cover 7, in another embodiment, one end of the sealing cover 7 is hinged on the side wall of the reagent box body 1, and the other end of the sealing cover 7 is connected by a snap fit to realize the closing of the cold-keeping bag placing cavity 6.

In order to facilitate storage of buffer solution reagent bottles, lysate reagent bottles and antibody reagent bottles more conveniently, in another embodiment, the top of the kit body 1 is further provided with a step wall 11, the first storage tank 3, the second storage tank 4 and the third storage tank 5 are all arranged below the step wall 11, and when the cover plate 2 is closed, the upper part of the step wall 11 is attached to the cover plate 2; the height of the step wall 11 is matched with the bottle cap thickness of the buffer solution reagent bottle, the lysate reagent bottle and the antibody reagent bottle. Therefore, when buffer solution reagent bottles, lysate reagent bottles and antibody reagent bottles are stored, the bottle caps of the buffer solution reagent bottles, the lysate reagent bottles and the antibody reagent bottles are closed, and the reagents in the buffer solution reagent bottles, the lysate reagent bottles and the antibody reagent bottles are prevented from being polluted.

In order to prevent the buffer solution reagent bottles, the lysate reagent bottles and the antibody reagent bottles from being damaged by vibration or impact during storage and transportation, in another embodiment, the bottoms of the first storage tank 3, the second storage tank 4 and the third storage tank 5 are provided with buffer pads.

In order to increase the storage stability of the reagent bottles, in another embodiment, the first storage groove 3, the second storage groove 4 and the third storage groove 5 are all arranged obliquely, and similarly, the buffer pads at the bottoms of the first storage groove 3, the second storage groove 4 and the third storage groove 5 are also arranged obliquely, so that the reagent bottles are placed in the corresponding storage grooves obliquely, and the stability of the reagent bottles can be enhanced well.

In order to facilitate taking the reagent kit body 1, in another embodiment, a clamping end 21 is disposed on one side of the cover plate 2, a first handle is disposed on a side surface of the reagent kit body 1, and a fastening end 12 matched with the clamping end 21 is disposed at the top of the first handle.

Since the liquid in the reagent bottle can cause pollution due to the inclined long-term basic bottle cap, in another embodiment, a second handle is further arranged on the reagent box body 1 on the opposite side of the first handle, and the first handle and the second handle are grabbed by two hands, so that the reagent box body 1 is convenient to take and move.

In order to improve the storage safety of the reagents in the reagent bottles, in another embodiment, white plastic reagent bottles are adopted for the buffer reagent bottles and the lysate reagent bottles, and labels corresponding to the buffer reagent bottles and the lysate reagent bottles are pasted outside the white plastic reagent bottles. The antibody reagent bottle adopts a brown glass reagent bottle, and a label corresponding to the lysate reagent bottle is adhered outside the brown glass reagent bottle.

In the specific use process, the cold-insulation bag is added into the cold-insulation bag placing cavity 6 to reduce the temperature inside the kit body 1 to 2-8 ℃, and then the buffer solution reagent bottle, the lysate reagent bottle and the antibody reagent bottle are respectively placed into the first storage tank 3, the second storage tank 4 and the third storage tank 5; wherein, the volume of the lysate reagent bottle is 80mL, and 50mL of erythrocyte lysate is filled in the lysate reagent bottle; a buffer solution reagent bottle with the volume of 150mL and filled with 110mL of buffer solution; the volume of the antibody reagent bottle is 1.5mL, and 0.8mL of antibody is filled in the antibody reagent bottle.

The TCR-T cell detection kit comprises the following steps:

(1) adding the antibody in an 8 mu L antibody reagent bottle into 50 mu L peripheral blood to be detected;

(2) incubating for 15 minutes at room temperature in dark;

(3) adding lysate in a lysate reagent bottle of 500 mu L;

(4) incubating for 15 minutes at room temperature in dark;

(5) adding the buffer solution in a 500-mu L buffer solution reagent bottle;

(6) standing for 10 minutes at room temperature in dark place;

(7) centrifuging at 400g for 5 minutes;

(8) discarding the supernatant, and adding 500. mu.L of buffer solution;

(9) repeating the centrifugation operation;

(10) discarding the supernatant, and adding 100 μ L buffer solution to resuspend the cell pellet;

(11) and (3) detecting APC and PE double positive cells on a flow type computer.

The peripheral blood TCR-T cell detection kit disclosed by the utility model directly marks TCR-T in peripheral blood by using two specific antibodies, so that the detection specificity is improved, the TCR-T cell proportion in the peripheral blood can be detected, the absolute number of TCR-T cells in the peripheral blood can also be detected, and the TCR-T cell detection time in the peripheral blood is shortened to 1 hour. Has the advantages of safe and convenient carrying and storage, simple and convenient operation, rapid result display, and the like.

The utility model is not limited to the above alternative embodiments, and any other various forms of products can be obtained by anyone in the light of the present invention, but any changes in shape or structure thereof, which fall within the scope of the present invention as defined in the claims, fall within the scope of the present invention.

Claims (9)

1. A TCR-T cell detection kit, characterized in that: the kit comprises a kit body, wherein a first storage groove for storing a buffer solution reagent bottle, a second storage groove for storing a lysate reagent bottle and a third storage groove for storing an antibody reagent bottle are arranged in the kit body; the kit body is characterized in that one side of the kit body is provided with a cold insulation bag placing cavity and a sealing cover for sealing the cold insulation bag placing cavity, and the cold insulation bag placing cavity is communicated with the first storage groove, the second storage groove and the third storage groove respectively in sequence.

2. A TCR-T cell detection kit as claimed in claim 1 wherein: and connecting channels are sequentially communicated among the first storage groove, the second storage groove and the third storage groove.

3. A TCR-T cell detection kit as claimed in claim 1 wherein: one end of the sealing cover is hinged to the side wall of the kit body, and the other end of the sealing cover is connected through a buckle to seal the cold-insulation bag placing cavity.

4. A TCR-T cell detection kit as claimed in claim 1 wherein: the top of the kit body is also provided with a step wall, the first storage groove, the second storage groove and the third storage groove are all arranged below the step wall, and when the cover plate is closed, the upper part of the step wall is attached to the cover plate; the height of the step wall is matched with the thicknesses of bottle caps of a buffer solution reagent bottle, a lysate reagent bottle and an antibody reagent bottle.

5. A TCR-T cell assay kit as claimed in claim 1 or 4, wherein: the bottoms of the first storage groove, the second storage groove and the third storage groove are all provided with cushion pads.

6. A TCR-T cell assay kit as claimed in claim 1 or 4, wherein: the first storage groove, the second storage groove and the third storage groove are all obliquely arranged.

7. A TCR-T cell detection kit as claimed in claim 1 wherein: a clamping end is arranged on one side of the cover plate, a first handle is arranged on the side face of the kit body, and a buckling end matched with the clamping end is arranged at the top of the first handle.

8. A TCR-T cell assay kit according to claim 7, wherein: a second handle is further arranged on the kit body and is positioned on the opposite side of the first handle.

9. A TCR-T cell detection kit as claimed in claim 1 wherein: the buffer solution reagent bottle and the lysate reagent bottle are both white reagent bottles, and labels corresponding to the buffer solution reagent bottle and the lysate reagent bottle are adhered outside the brown reagent bottle; the antibody reagent bottle is a brown glass reagent bottle.

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