CN113576556A - Oral mucosa cell sampling swab and sampling box - Google Patents
Oral mucosa cell sampling swab and sampling box Download PDFInfo
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- CN113576556A CN113576556A CN202111032215.8A CN202111032215A CN113576556A CN 113576556 A CN113576556 A CN 113576556A CN 202111032215 A CN202111032215 A CN 202111032215A CN 113576556 A CN113576556 A CN 113576556A
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- 238000005070 sampling Methods 0.000 title claims abstract description 109
- 210000002200 mouth mucosa Anatomy 0.000 title claims abstract description 53
- -1 Polyethylene Polymers 0.000 claims abstract description 19
- 239000004743 Polypropylene Substances 0.000 claims abstract description 15
- 229920001155 polypropylene Polymers 0.000 claims abstract description 15
- 239000004698 Polyethylene Substances 0.000 claims abstract description 14
- XECAHXYUAAWDEL-UHFFFAOYSA-N acrylonitrile butadiene styrene Chemical compound C=CC=C.C=CC#N.C=CC1=CC=CC=C1 XECAHXYUAAWDEL-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229920000122 acrylonitrile butadiene styrene Polymers 0.000 claims abstract description 14
- 239000004676 acrylonitrile butadiene styrene Substances 0.000 claims abstract description 14
- 229920000573 polyethylene Polymers 0.000 claims abstract description 14
- 235000017166 Bambusa arundinacea Nutrition 0.000 claims abstract description 9
- 235000017491 Bambusa tulda Nutrition 0.000 claims abstract description 9
- 241001330002 Bambuseae Species 0.000 claims abstract description 9
- 235000015334 Phyllostachys viridis Nutrition 0.000 claims abstract description 9
- 239000004793 Polystyrene Substances 0.000 claims abstract description 9
- 239000011425 bamboo Substances 0.000 claims abstract description 9
- 239000004800 polyvinyl chloride Substances 0.000 claims abstract description 9
- 239000002023 wood Substances 0.000 claims abstract description 9
- 239000003761 preservation solution Substances 0.000 claims description 22
- 238000007710 freezing Methods 0.000 claims description 13
- 230000008014 freezing Effects 0.000 claims description 13
- 238000005138 cryopreservation Methods 0.000 claims description 8
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 229920000915 polyvinyl chloride Polymers 0.000 claims description 4
- 210000000214 mouth Anatomy 0.000 abstract description 8
- 238000007790 scraping Methods 0.000 abstract description 5
- 210000004400 mucous membrane Anatomy 0.000 abstract 3
- 239000000243 solution Substances 0.000 description 14
- 210000003128 head Anatomy 0.000 description 12
- 239000007788 liquid Substances 0.000 description 9
- 238000004140 cleaning Methods 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 239000003755 preservative agent Substances 0.000 description 5
- 230000002335 preservative effect Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 235000002505 Centaurea nigra Nutrition 0.000 description 2
- 240000003323 Centaurea nigra Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000628997 Flos Species 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000003260 vortexing Methods 0.000 description 2
- 229920000742 Cotton Polymers 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
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- 238000003745 diagnosis Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 210000003813 thumb Anatomy 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B10/02—Instruments for taking cell samples or for biopsy
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
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Abstract
The invention provides an oral mucosa cell sampling swab and a sampling box, belonging to the technical field of medical instruments; the sampling swab comprises a sampling head and a handle; the sampling head and the handle are separable; the outer edge of the sampling head is in a sawtooth shape; the sampling head is provided with a groove; the sampling head is made of Polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), Polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), wood or bamboo. This application adopts the outward flange to get for scraping of the convenient oral cavity mucous membrane cell of sawtooth appearance, scrapes the oral cavity mucous membrane cell who gets to drop and falls on the recess of sampling head in, the collection of the oral cavity mucous membrane cell of being convenient for, but the volume of gathering is directly judged simultaneously. The oral mucosa cells collected by scraping with the oral mucosa cell sampling swab of the invention can not be adhered to the sampling head, thus being convenient for transferring the collected oral mucosa cells.
Description
Technical Field
The invention relates to the technical field of medical instruments, in particular to an oral mucosa cell sampling swab and a sampling box.
Background
Along with the increase of gene preservation, susceptible gene detection and diagnosis requirements in recent years, the demand of oral mucosa cell collection kits is also increased year by year. In the oral mucosa cell sampling process, the used collecting device directly influences the collecting amount, and further influences the follow-up research.
At present, the collection of oral mucosa cells is generally carried out by scraping by using a cotton swab or a swab with fluff. For example, CN 209404826U discloses an oral mucosal cell collection swab, which comprises a head and a holding part, wherein the head comprises a hard head and villi, and the villi is planted on the surface of the hard head and is used for collecting oral mucosal cells from the inner wall of the oral cavity. However, the swab head of the swab or the floss of the swab with the floss can adhere to a large amount of oral mucosa cells, and the transfer of the collected oral mucosa cells is not facilitated.
Disclosure of Invention
The invention aims to provide an oral mucosa cell sampling swab and a sampling box.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an oral mucosa cell sampling swab, which comprises a sampling head and a handle; the sampling device is characterized in that the outer edge of the sampling head is in a sawtooth shape;
the sampling head is provided with a groove;
the sampling head is made of polyethylene, polypropylene, polyvinyl chloride, polystyrene, acrylonitrile-butadiene-styrene copolymer, bamboo or wood.
Preferably, the deepest part of the groove is 0.4-0.6 cm.
Preferably, the width of the sampling head is smaller than the inner diameter of the cell freezing tube; the length of the sampling head is less than the height of the cell freezing tube.
Preferably, the length of the sampling head is 2-4 cm; the width of the sampling head is less than or equal to 1.1 cm.
Preferably, the joint of the handle and the sampling head is of a hollow structure.
Preferably, the material of the joint of the handle and the sampling head comprises polyethylene, polypropylene, polyvinyl chloride, polystyrene, acrylonitrile-butadiene-styrene copolymer, bamboo or wood.
Preferably, the handle is in the shape of a sheet; the length of handle is 8 ~ 10cm, the thickness of handle is 0.2 ~ 0.4 cm.
Preferably, the surface of the handle is engraved with corrugations; the corrugations are located in the middle of the handle.
The invention also provides an oral mucosa cell collecting box, which comprises the oral mucosa cell sampling swab, preservation solution and a cell freezing and storing tube in the scheme; the preservation solution comprises a DNA preservation solution and/or an RNA preservation solution.
Preferably, the specification of the cell freezing tube is 2mL or 5 mL.
The invention provides an oral mucosa cell sampling swab, which comprises a sampling head and a handle; the outer edge of the sampling head is in a sawtooth shape; the sampling head is provided with a groove; the sampling head is made of Polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), Polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), wood or bamboo. In the invention, the outer edge of the collecting head is in a sawtooth shape, so that the scraping of oral mucosa cells is facilitated, the scraped and fallen oral mucosa cells fall into the groove of the collecting head, the collection of the oral mucosa cells is facilitated, and the collecting amount can be directly judged. The material of the sampling head comprises Polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), Polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), wood or bamboo, and the collected oral mucosa cells are scraped and cannot be adhered to the sampling head, so that the collected oral mucosa cells are convenient to transfer.
Drawings
FIG. 1 is a front view of an oral mucosal cell sampling swab;
fig. 2 is a side view of an oral mucosal cell sampling swab.
Detailed Description
The invention provides an oral mucosa cell sampling swab, which comprises a sampling head and a handle; the outer edge of the sampling head is in a sawtooth shape; the sampling head is provided with a groove; the sampling head is made of Polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), Polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), bamboo or wood.
In the invention, the shape of the oral mucosa cell sampling swab is preferably spoon-shaped, more preferably straight spoon-shaped, so as to facilitate collection.
In the invention, the depth of the deepest part of the groove is preferably 0.4-0.6 cm, and more preferably 0.5 cm; the groove is convenient for the sampling swab to extend into the oral cavity for collecting and collecting the oral mucosa cells.
In the invention, the sampling head and the handle can be separated, so that the sampling head can be directly placed in the sample preservation solution.
In the invention, the width of the sampling head is smaller than the inner diameter of the cell freezing tube; the length of the sampling head is smaller than the height of the cell cryopreservation tube, and the sampling head separated from the handle can be directly placed into the cell cryopreservation tube so as to ensure that all the collected oral mucosa cells enter the cell cryopreservation tube. In the invention, the length of the sampling head is preferably 2-4 cm, and more preferably 3 cm; the width of the sampling head is preferably less than or equal to 1.1 cm; the thickness of the sampling head is preferably 0.2-0.4 cm, and more preferably 0.3 cm. A sampling head of this size was fitted to a 2mL cell cryopreservation tube.
In the invention, the connection part of the handle and the sampling head is preferably a hollow structure, so that the hollow structure can be conveniently broken under the action of external force, and the sampling head is separated from the handle; the hollow part is preferably circular in shape; the hollows are preferably arranged at intervals along the direction vertical to the handle; the number of the hollows is preferably 2-4, and more preferably 1. In the invention, the material of the joint of the handle and the sampling head comprises Polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), Polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), wood or bamboo.
In the present invention, the handle is shaped as a sheet; the length of the handle is preferably 8-10 cm, and more preferably 9 cm; the width of the handle is preferably 0.9-1.3 cm, and more preferably 1.1 cm; the thickness of the handle is preferably 0.2-0.4 cm, and more preferably 0.3 cm. A handle of this size is easy to hold.
In the invention, the total length of the oral mucosa cell sampling swab is preferably 10-14 cm, more preferably 11-13 cm, and most preferably 12 cm. This size is convenient for handling and manipulation.
In the invention, the surface of the handle is carved with ripples; the corrugations are located in the middle of the handle; the ripple has the effect of increasing the friction between the handle and the thumb and preventing the handle from sliding or sliding off to hurt the collected object in the process of scraping the oral mucosa.
The invention also provides an oral mucosa cell collecting box, which comprises the oral mucosa cell sampling swab, preservation solution and a cell freezing and storing tube in the scheme; the preservation solution comprises a DNA preservation solution and/or an RNA preservation solution.
In the invention, the specification of the cell freezing tube is 2mL or 5 mL.
The formulation of the DNA preservative solution and RNA preservative solution is not particularly limited, and conventional DNA preservative solutions and RNA preservative solutions in the art may be used.
In the invention, the use method of the oral mucosa cell collection box comprises the following steps:
adding DNA preservation solution or RNA preservation solution into the cell cryopreservation tube;
collecting oral mucosa cells by adopting the oral mucosa cell sampling swab;
after collection, the sampling head of the oral mucosa cell sampling swab is separated and put into a cell freezing tube filled with DNA preservation solution or RNA preservation solution.
In the invention, the oral mucosa cell collection mode is preferably to scrape the collected human oral cavity cheek.
In the present invention, the preservation temperature of the cell cryopreservation tube containing the sampling head in the above scheme is preferably-80 ℃.
In the invention, after a sampling head for separating oral mucosa cell sampling swabs is placed in a cell freezing tube containing DNA preservation solution, the method further comprises the steps of unfreezing for 20-40 min at 37 ℃, cleaning the oral mucosa cell sampling swabs, combining cleaning solution and the DNA preservation solution to obtain a combined solution, and extracting DNA of the combined solution. In the invention, the thawing time is preferably 25-35 min, and more preferably 30 min. In the invention, the reagent for cleaning the oral mucosa cell sampling swab comprises cell lysate; the purpose of cleaning the oral mucosa cell sampling swab is to avoid a small amount of DNA residue on the sampling swab. The method for extracting the DNA of the combined solution is not particularly limited, and the conventional method in the field can be adopted.
In the invention, after a sampling head of a sampling swab for separating oral mucosa cells is placed in a cell freezing tube containing RNA preservation solution, the sampling swab is unfrozen for 20-40 min at 37 ℃, the sampling swab for the oral mucosa cells is cleaned, cleaning solution and the RNA preservation solution are combined to obtain a combined solution, and RNA of the combined solution is extracted. In the invention, the thawing time is preferably 25-35 min, and more preferably 30 min. In the invention, the reagent for cleaning the oral mucosa cell sampling swab comprises cell lysate; the purpose of cleaning the oral mucosa cell sampling swab is to avoid a small amount of RNA residue on the sampling swab. The method for extracting RNA from the combined solution is not particularly limited, and the conventional method in the field can be adopted.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. 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.
In the embodiment, the front view and the side view of the oral mucosa cell sampling swab are respectively shown in fig. 1 and fig. 2, and the material of the oral mucosa cell sampling swab is preferably polypropylene and is integrally formed.
The collecting swab consists of a sampling head and a handle. The handle part is separable, and three round points are arranged at the joint of the sampling head and the handle in the drawing 1 and are of hollow structures. The total length of the collection swab is 12cm, the length of the collection head is 3.5cm, and the length of the handle is 8.5 cm. The collection swab had a width of 1.1cm and a thickness of 0.3 cm. Wherein the widest part of the groove of the sampling head is 1.1cm, and the deepest part is 0.5 cm. The handle is provided with a sculpture.
Example 1
1. The collected person rinses mouth with clear water, the buccal cavity mucosa cell sampling swab is adopted to gently scrape the buccal cavity cheek of the collected person, and the falling liquid is seen to fall in the groove of the collecting head. Breaking the hollow structure to separate the collecting head, putting the collecting head into a 2mL freezing storage tube containing DNA preservation solution (Tiangen, Beijing), storing at minus 80 ℃ for a long time, thawing the sample for 30min when DNA needs to be extracted, marking an EP tube in the period, and preparing a Kit (cargo number: 163050316) of QIAGEN DNAmicro Kit.
2. The thawed sample was gently removed from the collection swab with a small clean forceps, and the remaining fluid was added to a 2ml EP tube and centrifuged (6000g/5 min).
3. Carefully aspirate the liquid from step 2, leave the bottom pellet, leave a volume of about 30. mu.l, and discard the supernatant.
4. To 2ml of EP, 600. mu.l ATL and 600. mu. lAL were added, and 20. mu.l of Proteinase (mixed well before use) was added, followed by vigorous shaking for 10 seconds or more.
5. The sample in step 4 was placed in a 56 ℃ metal bath for 15min, during which time shaking was taken every 3 min.
6. Add 600. mu.l of absolute ethanol to a 2ml EP tube, mix by shaking immediately, vortex for 5s, shake off immediately, and stand for 5 min.
7. The adsorption column was removed, 700. mu.l of liquid was aspirated from the 2ml EP tube, carefully added to the column, allowed to stand for 1min, centrifuged at 16000g/2min, the lower layer of liquid was discarded, and the collection tube was replaced with a new one (this step was repeated twice, the first EP tube was aligned to the bearing (forward direction), and the second (reverse direction).
8. To the centrifuged column was added 500. mu. lAW1, 16000g/2min, and centrifuged (forward).
9. Abandoning the lower layer liquid in the step 9, replacing the collecting pipe, adding 500 mu lAW2, 16000g/2min, and centrifuging (reversing).
10. Abandoning the lower layer liquid in the step 10, replacing the collecting pipe, and idling and centrifuging for 16000g/3 min.
11. After idling, the column was air-dried for 5min, during which 2 sets of 1.5ml EP tubes were used.
12. After air-drying, the column was placed in a 1.5ml EP tube, 50. mu.l of TE was carefully added, and the column was subjected to a metal bath at 56 ℃ for 1min, left to stand at room temperature for 4min, and centrifuged (16000g/2 min).
13. And (6) performing detection on the machine.
Based on the first assay, step 13 was repeated after calculating the volume of TE added on the basis of ensuring that the second elution concentration reached 40 ng/. mu.l.
The results show that: the concentration of DNA detected on a computer is 136.8 ng/mul; the total amount is 6.84 mug, the method of the invention not only can effectively extract the DNA of the oral mucosa, but also can well complete the expected target.
Example 2
1. The collected person rinses mouth with clear water, uses the oral mucosa cell collecting swab to lightly scrape the oral cheek, and the liquid which is visible to fall off falls into the groove of the collecting head. The pick-up head was placed in a 2mL freezer containing RNA preservative fluid (Thermo, USA) and stored for a long period at-80 deg.C, the sample was thawed for 30min when DNA extraction was required.
2. Abandoning the sampling head, transferring the liquid into a 2ml EP tube, centrifuging for 5min at 12000 r and 4 ℃, abandoning the supernatant, leaving the precipitate, and draining the precipitate by absorbent paper.
3. Adding 1ml Trizol, blowing and uniformly mixing by using a gun head to avoid forming lumps, and vortexing for 2min until no obvious precipitate exists in the system.
4. Adding 200 μ l chloroform, shaking vigorously, vortexing for 45s, mixing and standing for 5 min.
5. 500. mu.l of the supernatant was taken and put into a 1.5ml EP tube and labeled.
6. Adding 500 μ l isopropanol, mixing, and standing at negative 20 deg.C for 2.5 hr.
7.13000g, and centrifuged at 4 ℃ for 15 min.
8. 500. mu.l of 75% ethanol was added to the supernatant.
9.13000g, centrifuged at 4 ℃ for 10 min.
10. Discarding the supernatant, centrifuging at 13000g for 5min at 4 ℃, sucking the liquid at the bottom of the tube, and drying in the air.
11. The precipitate was dissolved in 50. mu.l of enzyme-free water.
12. The metal bath is carried out at 60 ℃ for 2 min.
13. And (6) performing detection on the machine.
The results show that: the concentration of the detected RNA is 336.3 ng/mu l, the total amount is 16.81 mu g, the sampling sheet can effectively collect oral mucosa cells, and RNA can be well extracted.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. An oral mucosa cell sampling swab, the sampling swab comprises a sampling head and a handle; the sampling device is characterized in that the outer edge of the sampling head is in a sawtooth shape;
the sampling head is provided with a groove;
the sampling head is made of polyethylene, polypropylene, polyvinyl chloride, polystyrene, acrylonitrile-butadiene-styrene copolymer, bamboo or wood.
2. The oral mucosal cell sampling swab as recited in claim 1, wherein the deepest portion of the recess is 0.4 to 0.6cm deep.
3. The oral mucosal cell sampling swab as recited in claim 1, wherein the width of the sampling head is less than the inner diameter of the cell cryopreservation tube; the length of the sampling head is less than the height of the cell freezing tube.
4. The oral mucosal cell sampling swab as recited in claim 1 or 3, wherein the length of the sampling head is 2-4 cm; the width of the sampling head is less than or equal to 1.1 cm.
5. The oral mucosa cell sampling swab as recited in claim 1, wherein the handle is hollow at the connection with the sampling head.
6. The oral mucosal cell sampling swab as recited in claim 1, wherein the handle and the sampling head are made of polyethylene, polypropylene, polyvinyl chloride, polystyrene, acrylonitrile-butadiene-styrene copolymer, bamboo, or wood.
7. The oral mucosal cell sampling swab as recited in claim 1, wherein the handle is sheet-like in shape; the length of handle is 8 ~ 10cm, the thickness of handle is 0.2 ~ 0.4 cm.
8. A sampling swab according to claim 7, wherein the surface of the handle is corrugated; the corrugations are located in the middle of the handle.
9. An oral mucosa cell collecting box, comprising the oral mucosa cell sampling swab as claimed in any one of claims 1 to 8, a preservation solution and a cell freezing tube; the preservation solution comprises a DNA preservation solution and/or an RNA preservation solution.
10. The oral mucosa cell collection box of claim 9, wherein the size of the cell cryopreservation tube is 2mL or 5 mL.
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