CN214654868U

CN214654868U - Circulating tumor cell detection kit

Info

Publication number: CN214654868U
Application number: CN202023094695.4U
Authority: CN
Inventors: 张之胜; 张为国; 朱晓强; 邓晓洲; 史浩飞
Original assignee: Chongqing Institute of Green and Intelligent Technology of CAS
Current assignee: Chongqing Institute of Green and Intelligent Technology of CAS
Priority date: 2020-12-21
Filing date: 2020-12-21
Publication date: 2021-11-09
Anticipated expiration: 2030-12-21

Abstract

The utility model relates to a biology and medical detection field especially relates to a circulating tumor cell detect reagent box. Comprises a box body, a micro-through hole filter chip arranged in the box body, an injector, a filter, a reagent bottle and a reagent in the reagent bottle. The injector and the filter form a filter device, and a micro through hole filter chip is arranged in the filter and used for separating the circulating tumor cells. The shape of the micro through hole in the micro through hole filtering chip is a wheel-pendulum-shaped taper hole. The utility model discloses can effectively separate circulation tumor cell, avoid the cell jam, convenient to use detects with low costs.

Description

Circulating tumor cell detection kit

Technical Field

The utility model relates to a biology and medical detection field especially relates to a circulating tumor cell detect reagent box.

Background

Circulating tumor cells are tumor cells derived from primary or metastatic tumors that acquire the ability to detach from the basement membrane and invade blood vessels through the tissue matrix. Circulating tumor cells are increasingly becoming a hot research point for clinical liquid biopsy markers and are being widely used in clinic. Circulating tumor cells are present in very small amounts in peripheral blood, and may contain only 1 or a few, but about 1 million white blood cells and about 500 million red blood cells per 10ml of blood, and thus rapid and efficient detection of circulating tumor cells from peripheral blood is challenging.

The currently used circulating tumor cell detection methods mainly comprise two major types, namely immunochemical methods and physical methods. The immunochemical method mainly comprises an immunomagnetic bead method, and the CellSearch system in the United states is used for separating circulating tumor cells by an immunomagnetic bead technology, and has the defects of incapability of detecting tumor cells of non-epithelial sources or epithelial-mesenchymal transition and high analysis cost.

The physical methods include gradient centrifugation, microfluidic technology, membrane filtration, and the like. The disadvantages of the gradient centrifugation method are low separation efficiency, expensive equipment price and high analysis cost; microfluidic technology has high sensitivity but high analysis cost.

The membrane filtration method can obtain active tumor cells, but the conventional round hole has the problem of easy blockage. For example, the application numbers are: 201820906114.6, application date is: 2018.06.12, publication date is: 2018.12.28 discloses a 'detection kit for circulating tumor cells', which comprises a kit main body, a placing plate, an ice bag, a cooling box, heat-insulating cotton, a through hole, a containing box, a spring, an anti-skid sleeve, a test tube and a limiting block, wherein the placing plate is arranged at the lower side in the kit main body, the ice bag is arranged in the cooling box, the cooling box is arranged at the upper end surface of the placing plate, the heat-insulating cotton is arranged in the kit main body, the through hole is arranged at the upper side of the cooling box, the design solves the problem that the prior detection kit for circulating tumor cells has no cooling function, the containing box is fixed in the kit main body, the spring is arranged at the upper side in the containing box, the anti-skid sleeve is arranged at the upper side of the annular side of the test tube, the test tube is arranged at the upper side of the inner part of the containing box, and the limiting block is arranged at the annular side of the test tube, the design solves the problem that the positioning structure of the prior detection kit for circulating tumor cells is not enough, compared with the prior art, the utility model discloses the technical problem that actually solves is the easy problem of blockking up, the analysis is with high costs of pore membrane.

SUMMERY OF THE UTILITY MODEL

In view of this, the present invention provides a circulating tumor cell detection kit, which overcomes the problems of easy blockage of the membrane pores and high analysis cost in the prior art.

In order to achieve the above purpose, the utility model adopts the following technical scheme:

the utility model aims to provide a circulating tumor cell detection kit, which comprises a box body, wherein one end of the bottom of the box body is provided with an injector for injection, and also can be provided with other different instruments for applying pressure, and the other end of the bottom in the box body is provided with a filter containing a micro-through hole filter chip; a micro-through hole filtering chip for filtering is placed at one end of the upper part in the box body, reagent bottles are placed at the other end of the upper part in the box body, and different reagents are respectively placed in the reagent bottles; the micro-through hole filter chip and the reagent bottle are arranged on the upper sides of the injector and the filter. When the filtering device is required to be used, the filtering device can be used and normally works after being directly opened.

Furthermore, the micro-through hole array is composed of a plurality of micro-through holes with the aperture smaller than that of the circulating tumor cells, and the upper aperture of each micro-through hole is larger than the lower aperture. The upper hole comprises a first arc 221, a first line segment 222, a second arc 223 and a second line segment 224 which are sequentially connected end to end, and the lower hole comprises a third arc 226, a third line segment 227, a fourth arc 28 and a fourth line segment 229 which are sequentially connected end to end; the upper aperture is larger than the lower aperture, the upper aperture of the micro-through hole is larger than the lower aperture to form a cone, and the micro-through hole filter chip 21 is made of polymer materials, including PET, PC and PI.

Furthermore, the micro through holes are in a wheel pendulum shape and conical shape, and the micro through holes can be in other shapes, so that the requirement that the upper aperture of each micro through hole is larger than the lower aperture is met.

Further, the annular positioning frame includes: the clamping groove is formed between the inner frame and the outer frame.

Further, the injector comprises a cylinder body, wherein a pressure applying push rod with a piston is arranged in the cylinder body, and an inlet and an outlet are arranged at the lower end part of the cylinder body; and the inlet and outlet at the lower end part of the cylinder body are inserted into the injection inlet of the filter and connected. When the test is needed, the injector sucks the blood after cracking into the injector through the inlet and outlet, then the injector is connected with the filter to inject the blood into the filter through the pressure of the push rod, the filtration is carried out on the micro-through hole filter chip in the filter, and the circulating tumor cells are remained on the micro-through hole filter chip.

Further, the filter comprises an instrument lower cover with a buckle at the edge, and an outlet is arranged in the center of the instrument lower cover; a chip support frame with a support plate is arranged on the lower cover of the instrument; a micro through hole filter chip is arranged on the chip support frame; finally, the micro-through hole filter chip is covered by an instrument upper cover with an injection inlet; the injection inlet of the filter is connected with the inlet and outlet at the lower end part of the injector.

Furthermore, a sealing ring for sealing a space is arranged between the filter chip and the upper cover of the instrument, and the space between the filter chip and the upper cover of the instrument is a filtering middle position, so that the cleanness of the space is particularly important, and the sealing ring is used for sealing and connecting to prevent gaps.

Further, the reagent includes erythrocyte lysate, Phosphate Buffered Saline (PBS), paraformaldehyde, DAPI stain, etc., and may be substituted when other reagents are satisfied.

The beneficial effects of the utility model reside in that: provides a complete set of solutions for the isolation of circulating tumor cells. The micro-through holes are wheel-pendulum-shaped taper holes, so that the impact of fluid, the reaction of the hole walls to cells, the self-gravity of the cells and the like on the cells can be effectively weakened, the cells can roll on the surface of the holes in a self-adaptive manner, the blockage caused by single aperture size when the cells pass through is avoided, and the cell passing rate is improved. The filter device consisting of the injector and the filter is adopted, manual filtration is realized, and the use is convenient. The upper cover and the lower cover of the filter are fixed in a buckling mode, so that the filter is convenient to disassemble. The micro-through hole filter chip is made of polymer materials and is low in cost.

Drawings

FIG. 1 is a schematic view of the kit for detecting circulating tumor cells according to the present invention;

FIG. 2 is a schematic structural view of a micro-via filter chip of the present invention;

FIG. 3 is a schematic view of the wheel pendulum-shaped taper hole structure of the present invention;

FIG. 4 is a schematic view of the filter apparatus of the present invention;

fig. 5 is a schematic diagram of the filter structure of the present invention.

Reference numerals: the kit comprises a kit body 1, a micro-through hole 2 filtering chip, a syringe 3, a filter 4, a reagent bottle 5, a reagent 6, a chip carrier 21, a micro-through hole 22, a positioning frame 23, an inner frame 24, an outer frame 25, a first arc 221, a first line segment 222, a second arc 223, a second line segment 224, a side surface 225, a third arc 226, a third line segment 227, a fourth arc 228, a fourth line segment 229, a push rod 31, a piston 32, a cylinder 33, an inlet and outlet 34, an injection inlet 41, an injection inlet 42, an instrument upper cover, a 43 sealing ring, a chip supporting frame 44, an instrument lower cover 45, an outlet 46 and a 47 buckle.

Detailed Description

The examples are given for the purpose of better illustration of the invention, but the content of the invention is not limited to the examples. Therefore, those skilled in the art should appreciate that they can readily use the disclosed conception and specific embodiments as a basis for modifying or modifying other embodiments without departing from the scope of the present invention.

Example 1

Referring to fig. 1, the circulating tumor cell detection kit of the present invention comprises a box body 1, wherein an injector 3 for injection is placed at one end of the bottom of the box body 1, and a filter 4 is placed at the other end of the bottom in the box body; a micro-through hole filter chip 2 for filtering is placed at one end of the upper part in the box body, a reagent bottle 5 is placed at the other end of the upper part in the box body, and different reagents 6 are respectively placed in the reagent bottles 5; the filter chip 2 and the reagent bottle 5 are placed on the upper side of the syringe 3 and the filter 4. The reagent 6 comprises erythrocyte lysate, Phosphate Buffer Solution (PBS), paraformaldehyde and DAPI stain.

It should be noted that the micro-via array is composed of a plurality of micro-vias 22 with smaller apertures than the circulating tumor cells, and the upper aperture of the micro-vias 22 is larger than the lower aperture. The shape of the micro-via 22 also includes other shapes, the first line segment 222 and the second line segment 224 are common tangents of the first circular arc 221 and the second circular arc 223, the third line segment 226 and the fourth line segment 228 are common tangents of the third circular arc 227 and the fourth circular arc 229, so as to form a cone shape, and the annular positioning frame 223 includes: the inner frame 224 and the outer frame 225 form a clamping groove between the inner frame 224 and the outer frame 225.

Further, the injector 3 comprises a cylinder 33, a pressure applying push rod 31 with a piston 32 is arranged in the cylinder 33, and an inlet and an outlet 34 are arranged at the lower end of the cylinder 33; the filter 4 comprises an instrument lower cover 45 with a buckle 47 at the edge, and an outlet 46 is arranged in the center of the instrument lower cover 45; a chip support frame 44 with a support plate is arranged on the instrument lower cover 45; the chip support frame 44 is provided with a micro through hole filter chip 2; finally, the micro-through hole filter chip 2 is sealed by an instrument upper cover 42 with an injection inlet 41; the injection inlet of the filter 4 is connected with the inlet and outlet 33 at the lower end part of the injector 3. The inlet and outlet 33 of the injector 3 is inserted into the injection inlet 41 of the filter 4.

It should be noted that, a sealing ring 43 for sealing a space is disposed between the micro-via filter chip 2 and the instrument cover 42, and a spatial position between the micro-via filter chip 2 and the instrument cover 42 is a filtering intermediate position, so that the cleanliness of the spatial position is particularly important, and therefore, the sealing ring 43 is used for sealing connection to prevent a gap. The lower end of the cylinder 33 is provided with an inlet and outlet 34 which is inserted into the injection inlet 41 of the filter 4 for connection. When a test is required, the injector 3 sucks the lysed blood into the injector 3 through the inlet and outlet 34, then the filter 4 is connected to inject the blood into the filter 4 by pressing the push rod 31, the blood is filtered on the filter chip 2 in the filter 4, and the circulating tumor cells are retained on the micro-through hole filter chip 2.

Referring to fig. 1, the utility model includes: a circulating tumor cell detection kit comprises a kit body 1, a micro-through hole filter chip 2 arranged in the kit body 1, an injector 3, a filter 4, a reagent bottle 5 and a reagent 6 in the reagent bottle 5.

Referring to fig. 2, the micro via filter chip 2 is composed of a chip carrier 21, micro vias 22, and a positioning frame 23.

Referring to fig. 3, the micro through hole 22 is a pendulum-shaped taper hole, the pendulum-shaped taper hole is composed of an upper hole, a lower hole and a side surface, the upper hole is formed by sequentially connecting a first arc 221, a first line segment 222 and a second arc 223 end to end, and the first line segment 222 and the second line segment 224 are common tangents of the first arc 221 and the second arc 223. The lower hole is formed by sequentially connecting a third arc 226, a third line segment 227, a fourth arc 228 and a fourth line segment 229 end to end, and the third line segment 227 and the fourth line segment 229 are common tangents of the third arc 226 and the fourth arc 228. The upper hole is larger than the lower hole, and the upper hole, the lower hole and the side face form a wheel-pendulum-shaped taper hole together.

Referring to fig. 4, the syringe 3 is composed of a push rod 31, a piston 32, a cylinder 33, and a cylinder inlet/outlet 34. The syringe barrel port 34 is inserted just above the instrument top cap injection port 41.

Referring to fig. 5, the filter 4 is composed of an upper cover injection inlet 41, an upper cover 42, a sealing ring 43, the micro-via filter chip 2, a chip support 44, a lower cover 45 and a lower cover outlet 46. The upper instrument cover 42 and the lower instrument cover 45 are internally provided with a buckle 47, and the upper instrument cover 42 and the lower instrument cover 45 are fixed through the buckle 47.

Example 2

A few milliliters of blood of a tumor patient is put into a centrifuge tube, 3 times of volume of erythrocyte lysate is added, shaking is carried out repeatedly for 5 minutes to ensure that the erythrocytes are fully lysed, and then the lysed blood is sucked into an injector 3. The upper cover 42 of the instrument is opened, the sealing ring 43 is taken out, then the micro-through hole filter chip 2 is placed on the chip support frame 44, and then the upper cover 42 of the instrument and the lower cover 45 of the instrument are locked by a buckle. The barrel access port 34 of the syringe is then inserted just into the filter top cap injection port 41 and brought into intimate contact. Then push the push rod 31 by hand, squeeze and filter at uniform speed, the blood enters the injection inlet 41 of the filter upper cover from the inlet and outlet 34 of the cylinder, and then passes through the micro-through hole filter chip 2. The circulating tumor cells with larger volume are trapped on the micro-via filter chip 2, and the normal cells that have been lysed or have smaller volume pass through the micro-vias 22, and after the filtration is completed, they are washed 3 times with Phosphate Buffered Saline (PBS). And opening the filter 4, taking out the micro-through hole filter chip 2, and reserving tumor cells on the micro-through hole filter chip 2. Placed in Phosphate Buffered Saline (PBS). Then, the tumor cells on the micro-through hole filter chip can be desorbed by using the enzyme B for cell culture. Or through fixing, blocking, antibody incubation and staining, and finally detecting the tumor cell types and numbers under a darkroom microscope.

Finally, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the present invention can be modified or replaced by other means without departing from the spirit and scope of the present invention, which should be construed as limited only by the appended claims.

Claims

1. A circulating tumor cell detection kit is characterized by comprising a kit body (1), wherein one end of the bottom in the kit body (1) is provided with an injector (3) for injection, and the other end of the bottom in the kit body is provided with a filter (4); a micro-through hole filtering chip (2) for filtering is placed at one end of the upper part in the box body, a reagent bottle (5) is placed at the other end of the upper part in the box body, and different reagents (6) are respectively contained in the reagent bottles (5); the micro-through hole filter chip (2) and the reagent bottle (5) are arranged on the upper sides of the injector (3) and the filter (4).

2. The detection kit according to claim 1, wherein the micro-via filtration chip (2) comprises a chip carrier (21) and a micro-via array distributed on the chip carrier (21), the micro-via array is composed of a plurality of micro-vias (22) with a pore size smaller than that of the circulating tumor cells, and the upper pore size of the micro-vias (22) is larger than the lower pore size.

3. The test kit according to claim 2, wherein the micro-through-hole (22) is in the shape of a pendulum cone.

4. The test kit according to claim 2, wherein the chip carrier (21) is provided at its edge with a positioning frame (23) for positioning.

5. The test kit of claim 4, wherein the positioning frame comprises: an inner frame (24) and an outer frame (25).

6. The test kit according to claim 1, characterized in that the injector (3) comprises: the device comprises a cylinder (33), wherein a pressing push rod (31) with a piston (32) is arranged in the cylinder (33), and an inlet and an outlet (34) are arranged at the lower end of the cylinder.

7. The test kit according to claim 1, wherein the filter (4) comprises a lower cover (45) of the apparatus having a snap (47) at the edge, the lower cover (45) of the apparatus having an outlet (46) at the center; a chip support frame (44) with a support plate is arranged on the instrument lower cover (45); a micro through hole filter chip (2) is arranged on the chip support frame (44); finally, the micro-through hole filter chip (2) is covered by an instrument upper cover (42) with an injection inlet (41); the injection inlet (41) of the filter (4) is connected with the inlet and outlet (34) at the lower end part of the injector (3), and the inlet and outlet (34) at the lower end part of the cylinder body (33) of the injector (3) is inserted into the injection inlet (41) of the filter (4) to be connected.

8. The test kit according to claim 7, wherein a sealing ring (43) for sealing a space is provided between the micro-via filter chip (2) and the instrument top cover (42).