CN118025635A - Sample collection and processing device and application thereof - Google Patents

Abstract

The invention discloses a sample collection and processing device and application thereof, comprising a collection container, a storage container, a sample collection device, a plunger type filter and a sample preservation tube; the top of the collecting container is provided with an opening, and the bottom of the collecting container is provided with a first diversion hole and a second diversion hole with different heights; the sample collection device comprises a tubular structure, a piston, a push rod, a bottom diversion hole axially arranged in a columnar mode, a bottom transverse duct radially arranged in a columnar mode and a water outlet; the plunger filter is matched with the bottom transverse pore canal; the sample collection device is arranged at one side of the collection container and is used for collecting a sample flowing into the collection container; the storage container is arranged below the collection container and is used for storing redundant samples after the sample collection and treatment device is used for collecting the samples. The device can independently complete the whole process from sample collection to cell preservation treatment by non-professional staff without devices such as a centrifugal machine. The method is simple to operate, saves time and labor, and can obtain the cell or nucleic acid sample within 5-10 minutes.

Description

Sample collection and processing device and application thereof

Technical Field

The invention belongs to the field of biomedical treatment, relates to sample collection consumables, and particularly relates to a sample collection processing device which is convenient to operate and prevents pollution and application thereof.

Background

POCT (Point-of-CARE TESTING) is an instant inspection mode, and has the advantages of rapid detection, field sampling, simple and convenient operation and the like compared with the traditional inspection mode, so that the POCT has great significance in the fields of public health prevention and control, chronic disease monitoring, home self-inspection and the like. Especially in the field of home self-inspection, POCT is increasingly paid attention to because of the advantages of small size, low cost, simple and convenient operation and rapid detection.

Along with the popularization and development of various physical examination screening technologies, the acceptance of masses to various sample sampling means is higher and higher, but partial screening means are often influenced by privacy, religious beliefs, social customs and the like and are conflicted by users, so that the popularization difficulty is increased, such as traditional cervical cancer screening, AIDS screening means and the like. Studies show that the exfoliated cells contained in the early stage urine of morning urine can reflect the cytopathic degree of the lower genital tract to a certain extent and have higher sensitivity, so that cervical swabs can be replaced to a certain extent as a conventional screening means. And because of the low invasiveness of urine detection, urine detection has higher acceptance degree compared with cervical swabs, thereby being more beneficial to popularization of screening. Currently, urine detection is mostly used for self-detection of cervical cancer and AIDS.

The existing urine cell collecting device is complex in structure and inconvenient to operate, cells are often required to be collected and treated by a centrifuge and other machines, and follow-up cell collection and treatment are carried out by professionals, so that popularization and popularization of home self-inspection are not facilitated. And these devices are all direct with urine save and transportation, need store the transportation a large amount of urine during transportation, need add anti-degradation reagent in urine, occupy a large amount of volumes in transportation preservation, follow-up processing is wasted time and energy.

Disclosure of Invention

In order to solve the problems, the invention provides a sample collecting and processing device which is beneficial to autonomous collection at home, convenient to operate, pollution-preventing, convenient to preprocess and store collected samples and capable of realizing efficient transportation.

It is a further object of the present invention to provide the use of the aforementioned sample collection and processing device for rapid collection, preservation and efficient transportation of biological samples for domestic and/or commercial use.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

The invention provides a sample collection and processing device, which comprises a collection container, a storage container, a sample collection device, a plunger type filter and a sample preservation tube, wherein the collection container is provided with a sample storage tube; the top of the collecting container is provided with an opening, and the bottom of the collecting container is provided with a first diversion hole and a second diversion hole with different heights; the sample collection device comprises a tubular structure, a piston, a push rod, a bottom diversion hole axially arranged in a columnar mode, a bottom transverse duct radially arranged in a columnar mode and a water outlet; the plunger filter is matched with the bottom transverse pore canal; the sample collection device is arranged on one side of the collection container and is used for collecting a sample flowing into the collection container; the storage container is arranged below the collection container, and is used for storing redundant samples after the sample collection processing device is used for collecting the samples.

In the invention, the sample collecting device is an open collecting device, the bottom is provided with two flow guide holes with different heights, liquid is respectively led into the storage container and the sample collecting device, the storage container can be placed on a plane, the upper part of the sample collecting device is provided with a piston and a push rod thereof, a radial cylindrical hole is arranged below a liquid outlet at the lower part of the sample collecting device for placing a plunger type filter, the plunger type filter can be inserted below the sample collecting device, the filter can be extracted and placed in a sample processing tube after the sample is trapped, the lower part of the sample processing tube is provided with a liquid outlet, and the liquid in the tube can be extruded from the lower part by extruding the tube body.

Samples collected by the device of the present invention include, but are not limited to, natural bodies of water, plant juices, animal body fluids, or excretions thereof, intended to efficiently collect and capture cellular debris therein, as an alternative use scenario, such as collecting animal urine ex vivo, and capturing cellular debris therein.

The external shape of the sample collection and processing device is not limited, and as a preferable mode of the present invention, the collection container has an open design fitting the human body to prevent splash. In order to prevent liquid from directly flowing away from the diversion holes on the upper side of the collection container, a slightly inclined baffle is arranged on the upper side of the diversion holes so as to ensure that the liquid flows into the sample collection device from the diversion holes on the lower side at the first time, and the redundant liquid flows into the storage container from the diversion holes on the upper side.

The storage container is a flat bottom big belly container, so that the storage container can be conveniently placed on a flat ground and has a volume of 500-1000mL. The structure is integral with the collection container and the sample acquisition device described above.

As a preferred aspect of the present invention, the lower edge of the first deflector hole is higher than the upper edge of the second deflector hole, and the first deflector hole communicates with the storage container; the second deflector aperture is in communication with the sample acquisition device.

As a preferable mode of the invention, the upper edge of the first diversion hole is provided with a baffle plate.

As a preferable mode of the invention, the upper part of the storage container, the collecting container above the storage container and the sample collecting device are integrated, and a handle is arranged on one side of the collecting container far away from the sample collecting and processing device.

In the present invention, the collection container is tapered or ox horn-like in shape to prevent cell debris in the sample from hanging up. The volume of the collection vessel is preferably 50-1000mL, more preferably 300-500mL. The collecting container is higher on one side of the handle, so that the liquid in the collecting container can not splash out, and therefore a user can not touch a sample in the operation process, and pollution is avoided. The outer side wall of the collecting container is provided with a handle which is respectively connected with the collecting container and the storing container.

As a preferable scheme of the invention, the sample collection device further comprises a pressing piece, one end of the pushing rod is fixed with the pressing piece, the other end of the pushing rod is fixed with the piston, and the lower part of the piston is an inverted round table and is matched with the shape of the lower part of the tube body; the initial position of the piston is arranged at the top of the sample collection device, the bottom of the tube body structure gradually contracts to a bottom diversion hole, the tube body structure is of a conical structure, and a hole is formed in the center of the circle to divert liquid; the bottom transverse pore canal is arranged below the bottom diversion hole, and the diameter of the bottom transverse pore canal is larger than that of the bottom diversion hole and is used for inserting the plunger filter.

In the present invention, the volume of the sample collection device is 10 to 100mL, more preferably 20 to 50mL, as required. In the initial condition, the piston is positioned at the top, and is connected with a push rod, so that the piston can be pushed down to the bottom of the pipe by pushing the push rod externally. In the initial state, an opening is arranged on the inner side of the top of the pipe wall below the piston, and the opening is the second diversion hole of the collecting container. The bottom transverse duct aperture below the outlet is slightly larger than the bottom deflector aperture to house the plunger filter.

As a preferable scheme of the invention, the plunger filter comprises a plunger, an embedded filter membrane and a bottle cap; the plunger is cylindrical, a through hole with the same diameter is arranged at a position corresponding to the bottom diversion hole on the plunger, and a rigid embedded filter membrane is arranged in the through hole; the end of the plunger is a bottle cap.

In the invention, the diameter of the cylindrical plunger is consistent with that of the bottom transverse duct at the bottommost part of the sample collecting device, after the cylindrical plunger is inserted, a through hole with the same diameter is arranged at the corresponding position of the bottom diversion hole of the sample collecting device, and an embedded filter membrane is loaded in the through hole, and can intercept solid substances in a liquid sample, including various cells, tissue components and the like. The filtering membrane comprises PES membrane, glass fiber, microporous filter core and the like, and different filters can be selected based on different liquid environments.

As a preferable scheme of the invention, the sample preservation tube comprises a tube body and a spiral cover, wherein the upper part of the tube body is a cylinder, the lower part of the tube body is a cone, an upper opening is matched with the sample collection device, and a bulge is arranged at the bottom of the lower cone; the spiral cover is internally provided with a cavity for accommodating the lower part of the pipe body, and the cavity is provided with a circular gasket matched with the protrusion.

In the invention, the upper part of the sample preservation tube is a cylinder, the bottom is a reverse cone, the total volume is 5-10mL, and the tube body is made of transparent soft plastic. The upper part is provided with threads which are matched with the bottle cap of the plunger filter, thereby sealing the bottle body. The bottom of the lower cone is provided with a liquid outlet and a matched bottle cap, the inside of the bottle cap is a cone matched with the bottle bottom, the outside of the bottle cap is a cylinder convenient to operate, and the outer ring of the bottle cap is provided with anti-skid patterns. A rubber ring is arranged at the bottom of the bottle cap cone to prevent liquid in the tube from overflowing. The plunger filter is placed in a pipe, the bottle cap is screwed, the liquid in the pipe is kept stand after shaking, after the cell is broken, the lower bottle cap is unscrewed, and the pipe wall is slightly extruded, so that the liquid in the pipe can be extruded.

As a preferable mode of the present invention, the sample-holding tube is provided with a sample-processing solution.

In the present invention, the sample processing fluid includes, but is not limited to, a cell preservation fluid and/or a nucleic acid releasing agent. In particular, a sample-holding tube for holding a cell preservation solution can be used for holding sample cells and long-distance transport; the sample preservation tube for storing the nucleic acid releasing agent can obtain a nucleic acid sample through pretreatment so as to facilitate subsequent nucleic acid detection, is convenient and quick, and can be suitable for various POCT instruments or test strips and the like.

The invention also provides application of the sample collection and treatment device in enrichment treatment of cell debris in urine and/or rapid collection of biological samples for commercial use, preservation and efficient transportation.

Compared with the prior art, the invention has the following beneficial effects:

The sample collection and processing device and the sample collection assembly provided by the invention can independently complete the whole process from sample collection to cell preservation by non-professional persons under the condition of no equipment such as a centrifugal machine and the like. The operation is simple, time and labor are saved, and the sample cells can be obtained within 5-10 minutes. The sample preservation tube contains a sample processing solution, which facilitates preservation or pretreatment of the sample cells, and is convenient to transport due to its small size.

Drawings

FIG. 1 is a schematic view showing the structure of a sample collection and treatment apparatus according to example 1.

Fig. 2 is a schematic view of the structure of the bottom channel of the sample acquiring device according to example 1.

Fig. 3 is a schematic structural view of the plunger filter 140 of embodiment 1.

Fig. 4 is a schematic view showing the use state of the sample-holding tube 150 of example 2.

FIG. 5 is a detection result of urine cell nucleic acid samples combined with LAMP technology in example 3.

In the figure, 110. Collecting container; 111. a handle; 112. a first deflector aperture; 113. a second deflector aperture; 114. a baffle; 120. a storage container; 130. a sample collection device; 131. a pressing member; 132. a push rod; 133. a piston; 134. a bottom deflector aperture; 135. a bottom transverse duct; 136. a water outlet; 140. a plunger filter; 141. a bottle cap; 142. a plunger; 143. a through hole; 144. a filtering membrane; 150. a sample storage tube; 151. a tube body; 152. screwing the cover; 153. a protrusion; 154. a circular gasket.

Detailed Description

In order to facilitate understanding of the technical means, the creation characteristics, the achievement of the objects and the effects achieved by the present invention, the present invention is further described below with reference to specific examples, but the following examples are only preferred examples of the present invention, not all of which are described in detail below. Based on the examples in the embodiments, those skilled in the art can obtain other examples without making any inventive effort, which fall within the scope of the invention. The experimental methods in the following examples are conventional methods unless otherwise specified, and materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.

Example 1

The embodiment provides a collecting device for collecting cells in urine by taking the urine as a sample. As shown in fig. 1, the device includes a collection container 110, a storage container 120, a sample acquisition device 130, and a plunger filter 140, wherein the plunger filter 140 is removable.

The collection container 110 is a transparent inverted cone-shaped funnel structure with a maximum volume of 300mL, and a handle 111 is arranged on the outer side of the collection container to be connected with the storage container 120 on the lower part, so that the whole sample collection and treatment device can be conveniently held and moved. In order to prevent cell debris in urine from hanging up, the inner wall of the collection container 110 should be smooth and seamless and prevent the liquid droplets from settling, and the collection container 110 has a certain resistance to deformation to prevent the urine from overflowing due to deformation. The bottom of the collecting container 110 has two first and second guide holes 112 (upper) and 113 (lower) having different heights, and the lower edge of the first guide hole 112 should be slightly higher than the upper edge of the second guide hole 113. Wherein the first deflector hole 112 may deflect the liquid into the lower storage container 120, and the second deflector hole 113 may deflect the liquid into the right sample acquiring device 130. It should be noted that the upper edge of the first deflector aperture 112 has a ring of baffles 114 to ensure that urine does not flow out of the first deflector aperture 112 at a first time and that urine preferentially flows to the second deflector aperture 113 and into the sample acquisition device 130. After the sample collection and processing device 130 fills with urine, excess urine flows from the first deflector aperture 112 to the lower reservoir 120.

The reservoir 120 is a flat bottom rigid barrel-like container with a volume of 1000mL, with the upper rim collapsing inwardly to support the upper collection vessel 110 and sample collection device 130. The outer wall of the storage container 120 is connected to the upper collection container 110 by a handle 111.

The sample collection device 130 generally comprises a tubular outer wall and moving means (push member 131 and push rod 132), piston 133 (rubber piston), bottom deflector orifice 134, bottom transverse channel 135 and water outlet 136. From top to bottom, the pressing piece 131, the push rod 132 and the piston 133 are combined into a whole, so that the liquid in the pipe can flow downwards through the pressing piece 131 and the push rod 132. The piston 133 is matched with the inner diameter of the pipe wall, and in the initial state, when the piston 133 is at the top, the pipe wall is provided with a hole communicated with the second diversion hole 113 (actually, the second diversion hole 113), and the upper edge is just flush with the lower edge of the piston 133. The bottom of the tubular inner wall gradually collapses to a bottom deflector hole 134 which passes up and down. In use, urine first enters the collection container 110 and flows into the sample acquisition device 130 from the second flow director aperture 113, and after the sample acquisition device 130 is full of urine, excess urine flows into the storage container 120 from the first flow director aperture 112. In addition, the middle part of the bottom diversion hole 134 is provided with a bottom transverse pore canal 135, the two pore canals are mutually perpendicular, and the pore diameter of the bottom transverse pore canal 135 is slightly larger than that of the bottom diversion hole 134, and the structure is shown in fig. 2. Wherein the bottom transverse bore 135 may be inserted with a removable plunger filter 140.

The plunger filter 140 is configured as shown in fig. 3. The body is a plunger 142 with a diameter matching the bottom transverse bore 135 in the sample acquisition device 130, which is secured to a threaded cap 141, the outside of the cap 141 being provided with anti-slip threads. The entire plunger filter 140 may be inserted into the bottom transverse bore 135 of the sample acquisition device 130. After insertion, the plunger 142 has a through hole 143 having the same diameter at a position corresponding to the bottom deflector hole 134, and a filtering membrane 144 is fixed to the middle of the through hole 143. The plunger filter 140 cooperates with the sample collection device 130 to provide fluid filtration and solids collection functions in urine. After the urine is collected, the whole urine collecting device is slightly inclined, residual urine is discharged from the first diversion hole 112, the pressing piece 131 is pressed downwards, the urine is driven to pass through the plunger filter 140 at the lower part, and solid residues in the urine are trapped on the filter membrane 144 arranged in the filter.

Example 2

The present embodiment provides a sample collection and processing device comprising the collection container 110, the storage container 120, the sample collection device 130, and the plunger filter 140 described in embodiment 1, and further comprising a sample holding tube 150 as shown in fig. 4. These products can be packaged in an assembled or disassembled form into products.

Referring to fig. 4, the sample-holding tube 150 includes a tube body 151 screwed with a screw cap 152 at the bottom, and has a total volume of 4mL, and the tube body 151 is made of transparent plastic material, and has a certain elasticity, so that the solution can be extruded conveniently. The upper portion of sample-holding tube 150 may be cooperatively sealed with vial cap 141 of plunger filter 140, thereby placing plunger 141 and filter membrane 144 portions of plunger filter 140 in sample-holding tube 150. The bottom of the tube body 151 of the sample-holding tube 150 is tapered, and the tip of the tapered bottom has a protrusion 153, and the protrusion 153 is penetrating, so that the liquid in the tube can be released by squeezing the tube body 151. In order to prevent leakage of the liquid in the tube during storage and to allow the sample-holding tube 150 to stand upright, the bottom of the sample-holding tube 150 has a screw cap 152 which is screw-engaged therewith. The outer ring of the screw cap 152 is cylindrical, so that the whole sample-holding tube 150 is conveniently erected, and the inside is tapered to be matched with the lower structure of the tube body. A circular gasket 154 is provided at the inner bottom end of the screw cap 152 at a position abutting against the lower end opening of the tube body 151 to better close the tube body 151 and prevent the liquid from overflowing from the lower end.

In use, after the collection device is disposed of, solid matter, such as tissue, from the urine is retained on the filter membrane 144 of the plunger filter 140. The plunger filter 140 is pulled out of the sample collection device 130 and placed in the sample holding tube 150 where the cell holding fluid or lysate is previously added. After screwing, the shaking is repeatedly reversed to detach the cell tissue on the filtering membrane 144 and release the nucleic acid sample therein. Then the lower end of the cap 152 is screwed off, and the nucleic acid sample can be released by lightly pressing the tube 151.

Example 3

Based on the embodiment 2, the embodiment uses the sample collecting component to collect urine cell nucleic acid samples, the detection site is a section of exon sequence of actin beta-actin of common internal reference gene of human cells, and the detection is carried out by combining with loop-mediated isothermal amplification (LAMP) technology. The sample preservation tube is pre-filled with a rapid nucleic acid releasing agent, and the rapid nucleic acid releasing agent comprises the following components: betaine 200mM,Tris 300mM,Tween 20 2%, potassium chloride 200mM, dithiothreitol (DTT) 0.1%, N-acetylcysteine 0.1%.

Morning urine was collected as provided in example 2, the filtered plunger filter was inserted into a sample holding tube containing the rapid nucleic acid releasing agent, the filter membrane was immersed in the nucleic acid releasing agent, the bottle cap was screwed on and left standing for 5 minutes, and the middle was suitably inverted and homogenized.

After the mixture is inverted and evenly mixed, the lower spiral cover is screwed off, the wall of the preservation pipe is extruded to release the nucleic acid sample, the nucleic acid sample is added into a loop-mediated isothermal amplification (LAMP) system, and after the mixture is evenly mixed, the mixture is placed into a fluorescent quantitative PCR instrument for isothermal amplification and fluorescent curve detection, and the procedure is carried out at 65 ℃ for 30 minutes. Judging whether the result is positive according to the fluorescence detection curve.

The complete DNA sequence for beta-actin was obtained from NCBI (National Center for Biotechnology Information ), and Primer design used version V5 of the Primer Explorer website.

In this example, the primer/probe sequence information used for LAMP is shown in table 1:

TABLE 1 primer/probe names

The LAMP system is shown in table 2:

TABLE 2 LAMP System

Composition of the components	Volume of
		Actin-F3(100μM)	0.2μL
Actin-B3(100μM)	0.2μL
		Actin-FIP(100μM)	1.6μL
Actin-BIP(100μM)	1.6μL
		Actin-LF(100μM)	0.4μL
Actin-LR-FAM(100μM)	0.1μL
		2×LAMP Mix	12.5μL
Nucleic acid sample	7.5μL
		ddH2O	0.9μL
Total	25μL

The fluorescence PCR instrument used was Applied Biosystems QuantStudio, the reaction procedure was 65℃for 30 minutes, the fluorescence channel was FAM, and fluorescence was detected every minute.

In this example, four test persons were used to test their urine and physiological saline was used as a negative control. The detection results are shown in Table 3, and the fluorescence curves are shown in FIG. 5:

TABLE 3 detection results

In this example, all tested human samples were positive, and all saline samples were negative, consistent with the expected results. The result shows that the device for collecting the cell in the urine can effectively intercept the cell debris sample in the urine and meet the requirement of nucleic acid loading in subsequent nucleic acid detection.

In the embodiment, the cell in urine is collected and transferred to the sample collecting tube only for 2-3 minutes, the nucleic acid is released only for 5 minutes, the whole process can obtain the nucleic acid quantity which is enough for the subsequent nucleic acid detection in less than 10 minutes, the operation is simple and convenient, the auxiliary devices such as a centrifugal machine and the like are not needed, and the whole process from sampling to result judgment can be completed in 1 hour by combining the technologies such as the subsequent isothermal amplification and the like.

It should be noted that the above embodiments are only for illustrating the application method of the present patent, and not all application ranges are described, and the detection method combined with isothermal amplification as described in the present embodiment is intended to illustrate the specific application scenario of the cell sample provided by the present invention, but the application of the present invention should not be limited to the LAMP technology. It should be noted that the nucleic acid sample processed by the device for collecting cells in urine provided by the invention can be widely applied to various nucleic acid detection technologies, including conventional PCR technology, constant temperature detection technology and the like. In addition, it is within the purview of the present patent to have modifications and variations based on the appearance and materials of the patent as claimed by the present patent for those skilled in the art.

Claims (9)

1. The sample collection and treatment device is characterized by comprising a collection container, a storage container, a sample collection device, a plunger type filter and a sample preservation tube; the top of the collecting container is provided with an opening, and the bottom of the collecting container is provided with a first diversion hole and a second diversion hole with different heights; the sample collection device comprises a tubular structure, a piston, a push rod, a bottom diversion hole axially arranged in a columnar mode, a bottom transverse duct radially arranged in a columnar mode and a water outlet; the plunger filter is matched with the bottom transverse pore canal; the sample collection device is arranged on one side of the collection container and is used for collecting a sample flowing into the collection container; the storage container is arranged below the collection container, and is used for storing redundant samples after the sample collection processing device is used for collecting the samples.

2. The sample collection and processing device of claim 1, wherein a lower edge of the first flow director aperture is higher than an upper edge of the second flow director aperture, the first flow director aperture in communication with the reservoir; the second deflector aperture is in communication with the sample acquisition device.

3. The sample collection and processing device of claim 1, wherein the upper edge of the first deflector aperture is provided with a baffle.

4. A sample collection and processing device according to claim 1, wherein the upper portion of the reservoir is integral with the overlying collection container and sample collection device, and a handle is provided on the side of the collection container remote from the sample collection and processing device.

5. The sample collection and processing device according to claim 1, further comprising a pressing member, wherein one end of the pushing rod is fixed to the pressing member, the other end of the pushing rod is fixed to the piston, and the lower portion of the piston is an inverted truncated cone and is matched with the lower portion of the tube body in shape; the initial position of the piston is arranged at the top of the sample collection device, the bottom of the tube body structure gradually contracts to a bottom diversion hole, the tube body structure is of a conical structure, and a hole is formed in the center of the circle to divert liquid; the bottom transverse pore canal is arranged below the bottom diversion hole, and the diameter of the bottom transverse pore canal is larger than that of the bottom diversion hole and is used for inserting the plunger filter.

6. The sample collection device of claim 1, wherein the plunger filter comprises a plunger, an embedded filter membrane, and a vial cap; the plunger is cylindrical, a through hole with the same diameter is arranged at a position corresponding to the bottom diversion hole on the plunger, and a rigid embedded filter membrane is arranged in the through hole; the end of the plunger is a bottle cap.

7. The sample collection and processing device according to claim 1, wherein the sample storage tube comprises a tube body and a screw cap, the upper part of the tube body is a cylinder, the lower part of the tube body is a cone, an upper opening is matched with the sample collection device, and a lower cone bottom is provided with a bulge; the spiral cover is internally provided with a cavity for accommodating the lower part of the pipe body, and the cavity is provided with a circular gasket matched with the protrusion.

8. A sample collection and processing device according to claim 7, wherein the sample holding tube has a sample processing solution disposed therein.

9. Use of a sample collection and processing device according to any one of claims 1-8 for cell debris enrichment treatment in urine and/or for commercial rapid collection, preservation and efficient transportation of biological samples.