A body fluid collection, pretreatment and storage device and its use method

Abstract

The present invention discloses a body fluid collection, pretreatment and preservation device and a method for using the same. The device comprises: a sampling tube, a push cylinder sliding in the sampling tube, a filter assembly arranged at the bottom of the sampling tube, a collection tube connected to the filter assembly, and a biological sample protective agent; the filter assembly comprises: a porous polymer filter plate arranged inside the sampling tube, and a filter head connected to the lower end of the sampling tube; the device and method of the present invention can eliminate the interference of mucin, bacteria, cell debris, and oral insoluble matter / impurities at one time under low resistance, complete the initial purification process, and improve the stability of biological molecules in body fluid samples, especially metabolic molecules, under non-low temperature conditions.

Description

Technical Field

[0001] The present invention relates to the field of detection and testing tools, and in particular to a body fluid collection, pretreatment and storage device and a method for using the same. Background Art

[0002] Saliva is a viscous liquid secreted by multiple salivary glands in the mouth. In addition to water, its main components include a large number of proteins, peptides, endogenous metabolites and inorganic ions. The exfoliated cells in the mouth also contain human DNA. Changes in saliva components have been shown by many studies to be closely related to chronic diseases and cancer. Among them, amino acids, fatty acids, urea, glucose, choline and their derivatives in saliva can directly reflect the level of human amino acid metabolism, lipid metabolism, urine circulation, glycolysis and choline metabolism from the metabolic level, thereby further reflecting whether the human body suffers from metabolic diseases such as cancer, diabetes, chronic kidney disease, Alzheimer's disease, etc., so it can be used as a relatively sensitive biomarker. The detection of saliva is conducive to early screening, early diagnosis and early intervention of diseases. Among the many types of human body fluids, saliva is the easiest and most comfortable biological sample to collect. It is a completely non-invasive sampling with low professional requirements, which is conducive to improving patient compliance and reducing diagnosis and treatment pressure, so it is increasingly valued in clinical practice.

[0003] The reliability of the biological information reflected by saliva samples depends to a large extent on a reasonable and standardized sampling process and pretreatment and storage process. Because the components of saliva samples are complex, they not only contain the molecules mentioned above, but also contain more than 1,000 kinds of bacteria, whose metabolic processes will seriously affect the composition of endogenous metabolic molecules and peptides in saliva. Cell fragments, insoluble matter and impurities in the mouth are more likely to contaminate saliva samples. The presence of a large amount of mucin in saliva is difficult to remove by centrifugation and other operations, which will seriously affect the subsequent extraction and enrichment of other biological molecules. Especially when the subsequent detection object is a metabolic molecule, it is necessary to eliminate the interference of multiple substances such as bacteria, cell fragments, mucin, oral insoluble matter, etc. at the same time to obtain a clear, clean and stable effective saliva sample. The market urgently needs a saliva collection device that can be used for routine test samples collected at home and stored and transported at non-low temperature, which is easy to operate, efficient and stable, and can integrate collection, pretreatment and storage.

[0004] The most common saliva collection device on the market now is a collection tube with a funnel accessory. Generally, the stability of the saliva sample is improved by squeezing the built-in DNA protectant in the collection tube to mix it with the sample or adding a DNA protectant, such as the sampling devices mentioned in patents CN210347197U and CN209032444U. This type of device collects non-irritating saliva passively produced by the human body, but the saliva samples collected by this type of device are highly viscous and cannot eliminate the interference of bacteria, cell debris and mucin, and contain a lot of oral debris. The collected saliva is directly added to the DNA preservation solution to ensure the appropriate ratio, and it is not suitable for the stability control of small molecules such as metabolism and polypeptides. Patent CN206523325U discloses a saliva collection and preservation device containing a filter membrane, a filter membrane with a pore size of 200nm is attached to the bottom of the device, and the saliva sample in the collection and preservation device is squeezed to pass through the filter membrane. Although this method can remove bacteria and physical residues, it is not very practical because the viscous protein in the saliva is highly viscous and easily clogs the filter membrane.

[0005] Another large category of saliva collection devices on the market requires the use of sampling absorbent cotton or absorbents to collect saliva. The saliva collection device inserts cotton wool into the mouth to absorb a sufficient amount of saliva sample, and then places the cotton wool in a special centrifuge tube for high-speed centrifugation to obtain a clarified liquid sample. This method requires a centrifuge to operate, and is only suitable for unified processing in laboratories, and cannot be applied to scenarios without professional equipment. Patents such as US7618591B2, US7850922B2, and US8641642B2 all disclose another type of collection device that can obtain clarified saliva without centrifugation, that is, glass fiber material or other adsorbents with strong adsorption force are inserted into the mouth to adsorb saliva, and then the adsorbed liquid is squeezed into the collection tube with the help of pressure. This type of sampling device can adsorb mucin with the help of an adsorbent, but it also cannot eliminate the interference of bacteria. The collection devices disclosed in patents US10023903B2, CN206486520U, and CN209734017 add filter tubes or filter membranes on this basis, which can simultaneously eliminate the interference of mucin, bacteria, impurities, etc. However, such devices containing sampling adsorbents need to insert the sampling head into the oral cavity, which is prone to induce irritation reactions. Although they can meet the collection and detection of DNA, etc., they will interfere with the components of metabolic molecules, polypeptides and other molecules in the oral cavity, causing inconsistent results. In addition, small molecules will dissolve out of the adsorbent under the action of body fluid samples, thereby affecting the reliability of biological information.

[0006] Therefore, how to provide a saliva collection, pretreatment and storage device that can not only ensure that saliva secretion is not disturbed by the stimulation of external sampling cotton, etc., but also eliminate the interference of mucin, bacteria, cell fragments, oral insoluble matter / impurities in an integrated low-resistance manner, complete the initial purification process, and improve the stability of metabolic molecule groups under non-low-temperature conditions, has become an urgent problem to be solved by those skilled in the art; the present invention solves such a problem. Summary of the invention

[0007] In order to address the deficiencies of the prior art, the purpose of the present invention is to provide a body fluid collection, pretreatment and preservation device and a method of use thereof. The device and method of the present invention can eliminate the interference of mucin, bacteria, cell debris, oral insoluble matter / impurities at one time under low resistance conditions, complete the initial purification process, and improve the stability of biological molecules in body fluid samples, especially metabolic molecules under non-low-temperature conditions.

[0008] In order to achieve the above object, the present invention adopts the following technical solution:

[0009] A body fluid collection, pretreatment and storage device comprises: a sampling tube, a push cylinder sliding inside the sampling tube, a filter assembly arranged at the bottom of the sampling tube, a collection tube connected to the filter assembly, and a biological sample protective agent; the filter assembly comprises: a porous polymer filter plate arranged inside the sampling tube, and a filter head connected to the lower end of the sampling tube.

[0010] In the aforementioned body fluid collection, pretreatment and preservation device, the porous polymer filter plate is a hydrophobic porous polymer filter plate, the plane size of the hydrophobic porous polymer filter plate is the same as the inner diameter of the sampling tube, the thickness is 1-3mm, and the pore size is 10-100μm.

[0011] The aforementioned body fluid collection, pretreatment and storage device, the filter head includes: a filter head body, and a low-adsorption microporous filter membrane arranged in the filter head body; the pore size of the low-adsorption microporous filter membrane is 0.2-0.5μm.

[0012] The aforementioned body fluid collection, pretreatment and storage device, the collection tube includes: a collection tube body, a pointed bottom formed at the bottom of the collection tube body, a tube cover connected to the collection tube body, and scales formed on the outer wall of the collection tube body.

[0013] In the aforementioned body fluid collection, pretreatment and storage device, the collection tube body is connected to the sampling tube by threads.

[0014] In the aforementioned body fluid collection, pretreatment and storage device, the tube opening of the collection tube body is larger than the diameter of the liquid outlet of the filter head.

[0015] The aforementioned body fluid collection, pretreatment and storage device also includes: a sampling funnel movably connected to the sampling tube.

[0016] In the aforementioned body fluid collection, pretreatment and storage device, the connection methods between the sampling tube and the sampling funnel include: threaded connection and glued connection.

[0017] In the aforementioned body fluid collection, pretreatment and preservation device, the biological sample protective agent includes: methanol, ethanol, acetonitrile, acetone, and a mixture of methanol and acetonitrile.

[0018] In the aforementioned body fluid collection, pretreatment and preservation device, the volume ratio of the biological sample protective agent to the sample is 2:1-5:1.

[0019] A method for using the aforementioned body fluid collection, pretreatment and storage device,

[0020] Step 1: The subject holds the upper end of the body fluid sampling tube and collects the body fluid into the sampling funnel. The body fluid is retained on the porous polymer filter plate. The sampling volume is 0.5-1mL;

[0021] Step 2: Separate the sampling funnel from the sampling tube, hold the tail of the push tube, and push the head of the push tube directly into the sampling tube. The sample passes through the porous polymer filter plate and filter head and enters the collection tube.

[0022] Step 3, separating the collection tube from the sampling tube, thus obtaining the pre-treated body fluid sample, with a volume of 0.15-0.3 mL;

[0023] Step 4: Pour the biological sample preservative into the collection tube, cover the tube tightly, and mix the biological sample preservative and body fluid sample evenly.

[0024] The present invention is beneficial in that:

[0025] The body fluid collection, pretreatment and storage device provided by the present invention can collect non-irritating saliva samples without introducing sampling cotton and sampling adsorbents, eliminate the interference of mucin, bacteria, cell debris, oral insoluble matter / impurities at one time under low resistance, and complete the initial purification process, during which no centrifuge equipment and other laboratory professional equipment are required;

[0026] The body fluid collection device disclosed in the present invention can realize the collection of saliva samples with relatively stable volume, thereby improving the consistency of the collected saliva samples;

[0027] The device of the present invention can be fixed in one piece or can be disassembled by threaded connection, and occupies a small volume, and is easy to place, store and uniformly handle later;

[0028] The body fluid collection, pretreatment and storage device provided by the present invention is simple to operate and highly efficient, suitable for operation by non-professionals, and has scalability in terms of sample types and sample detection objects;

[0029] The body fluid samples collected by the body fluid collection device of the present invention do not need to be stored at low temperatures;

[0030] The body fluid collection device of the present invention has a filtering component with a step-by-step filtering function, which can filter out viscous proteins, cell debris, impurities, oral insoluble matter and bacteria at the same time, thereby reducing the storage conditions and maintaining stability for more than one week under non-low-temperature storage conditions, and is suitable for non-centralized sampling, pretreatment and storage. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 is a schematic structural diagram of an embodiment of a body fluid collection device of the present invention;

[0032] Figure 2 It is a structural schematic diagram of an embodiment of a sampling tube of the present invention;

[0033] Figure 3 is a metabolic spectrum of a saliva sample collected by the body fluid collection device of the present invention, wherein the abscissa is the mass-to-charge ratio (m / z), without unit, and the ordinate is the peak intensity, without unit;

[0034] Figure 4 : is a mass spectra comparison diagram of saliva samples collected by the body fluid collection device of the present invention and a conventional collection device (funnel with collection tube), A is a spectrum of 3 consecutive tubes of saliva samples collected by the funnel with collection tube, B is a spectrum of 3 consecutive tubes of saliva samples collected by the body fluid collection device of the present invention, the abscissa is mass-to-charge ratio (m / z), no unit, and the ordinate is peak intensity, no unit;

[0035] Figure 5 It is a comparison chart of mass spectra of subjects passing through body fluid collection devices containing hydrophilic porous polymer filter plates and hydrophobic porous polymer filter plates, A is the hydrophilic sieve plate, B is the hydrophobic sieve plate, the abscissa is the mass-to-charge ratio (m / z), no unit, and the ordinate is the peak intensity, no unit;

[0036] Figure 6 It is the mass spectrum of the saliva sample collected by the subject through the body fluid collection device without a filter plate but with a filter head containing a microporous filter membrane. The horizontal axis is the mass-to-charge ratio (m / z), without unit, and the vertical axis is the peak intensity, without unit;

[0037] Figure 7 is the coefficient of variation of the mass spectra of the saliva samples collected using the body fluid collection device of the present invention under three different storage conditions;

[0038] Figure 8 It is a representative spectrum and coefficient of variation of saliva samples collected by the body fluid collection device of the present invention at different time periods. The horizontal axis of the spectrum is the mass-to-charge ratio (m / z), which has no unit, and the vertical axis is the peak intensity, which has no unit.

[0039] Meaning of the reference numerals in the figures:

[0040] 1 sampling tube, 2 push tube, 21 head, 22 tail, 3 filter assembly, 31 porous polymer filter plate, 32 filter head, 321 low adsorption microporous filter membrane, 4 collection tube, 41 tube cover, 5 biological sample preservative, 6 sampling funnel. DETAILED DESCRIPTION

[0041] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0042] It should be noted that the body fluids of the present invention include body fluid samples such as saliva, sweat, urine, and sputum.

[0043] like Figure 1 The body fluid collection, pretreatment and preservation device shown includes: a sampling tube, a push cylinder sliding in the sampling tube, a filter assembly arranged at the bottom of the sampling tube, a collection tube connected to the filter assembly, and a biological sample preservative; the filter assembly includes: a porous polymer filter plate arranged inside the sampling tube, and a filter head connected to the lower end of the sampling tube.

[0044] The porous polymer filter plate can be a hydrophilic porous polymer filter plate or a hydrophobic porous polymer filter plate; Figure 2 As shown, as a preferred embodiment, the porous polymer filter plate uses a hydrophobic porous polymer filter plate, and the hydrophobic porous polymer filter plate is one of porous high molecular weight polyethylene and porous polymer polypropylene. The plane size of the hydrophobic porous polymer filter plate is the same as the inner diameter of the sampling tube, with a thickness of 1-3 mm and a pore size of 10-100 μm. It can intercept and adsorb mucin, cell debris, and insoluble matter in body fluid samples such as saliva. The liquid entering the sampling tube without pressure will not automatically pass through the porous polymer filter plate. When sampling, the body fluid sample is automatically intercepted above the porous polymer filter plate at the bottom of the sampling tube.

[0045] Compared with the hydrophilic porous polymer filter plate, the hydrophobic porous polymer filter plate can ensure that the liquid inside the sampling tube will not automatically pass through the porous polymer filter plate without external pressure. During sampling, the body fluid sample is automatically retained above the porous polymer filter plate at the bottom of the sampling tube, and the volume of the body fluid sample entering the sampling tube can be observed more intuitively. At the same time, the hydrophobic porous polymer filter plate has a low adsorption capacity for small molecule metabolites in body fluids, and can maximize the consistency of the sample after passing through the body fluid collection device.

[0046] like Figure 2 As shown, the filter head includes: a filter head body, and a low-adsorption microporous filter membrane disposed in the filter head body; the pore size of the low-adsorption microporous filter membrane is 0.2-0.5 μm, and can filter bacteria and sticky substances that are not filtered by the porous polymer filter plate.

[0047] The collecting tube includes: a collecting tube body, a pointed bottom formed at the bottom of the collecting tube body, a tube cover connected to the collecting tube body, and a scale formed on the outer wall of the collecting tube body. The collecting tube can accommodate a sample volume of 2 mL. The collecting tube body is connected to the sampling tube by a thread. The tube mouth of the collecting tube body is larger than the diameter of the liquid outlet of the filter head. After sampling, it is detached from the bottom of the sampling tube and can be sealed for storage. The scale can indicate the volume of the sample entering the sampling tube. The volume of the fluid sample passing through the filter assembly is basically constant. When the volume of the saliva sample entering the sampling tube is 0.5-1 mL, the volume of the clarified saliva sample collected in the collecting tube ranges from 0.15 to 0.3 mL, thereby realizing the collection of body fluid samples with relatively stable volume.

[0048] A body fluid collection, pretreatment and storage device also includes: a sampling funnel movably connected to a sampling tube. The connection between the sampling tube and the sampling funnel includes: threaded connection and glued connection. As a preferred embodiment, the sampling funnel is connected to the sampling tube thread by a thread, and the sampling funnel can be removed after sampling.

[0049] The head of the push tube is made of rubber, and its size is the same as the inner diameter of the sampling tube. The tail of the push tube is the hand-held part. When the push tube is pushed into the sampling tube, the air between the liquid at the bottom of the sampling tube and the head of the push tube is compressed, generating a pressure of 10-50N, causing the liquid at the bottom of the sampling tube to pass through the hydrophobic porous polymer filter plate and filter head into the collection tube.

[0050] The biological sample preservative is a reagent containing an organic solvent, which is stored separately in a well-sealed reagent bottle. The organic solvent includes: methanol, ethanol, acetonitrile, acetone, and a mixture of methanol and acetonitrile. As a preferred embodiment, the biological sample preservative is a mixture of methanol and acetonitrile, and the volume ratio of methanol to acetonitrile includes: 3:1, 1:1, 1:3.

[0051] Using the above device, saliva samples were obtained in the following manner.

[0052] The steps for using the saliva collection, pretreatment and storage device are as follows:

[0053] Step 1: After cleaning the oral cavity, the subject calmly produces saliva for 2-5 minutes, holds the upper end of the saliva sampling tube, and spits the saliva into the sampling funnel. The fluid enters the sampling tube through the sampling funnel and is intercepted on the porous polymer filter plate at the bottom of the sampling tube. The sampling volume is 0.5-1mL;

[0054] Step 2, separate the sampling funnel from the upper part of the sampling tube, hold the tail of the push tube, and push the head of the push tube directly into the sampling tube to compress the gas in the sampling tube. The body fluid in the main body of the sampling tube is subjected to the pressure from above and passes through the porous polymer filter plate and filter head of the filter assembly in sequence and enters the collection tube below the sampling tube;

[0055] Step 3, detach the collection tube from the bottom of the sampling tube to obtain the pre-treated saliva sample with a volume of 0.15-0.3 mL;

[0056] Step 4, pour the biological sample protective agent into the collection tube, cover the tube tightly, and shake vigorously to mix the protective agent and the saliva sample evenly. The biological sample protective agent includes: methanol, ethanol, acetonitrile, acetone, and a mixture of methanol and acetonitrile; the volume ratio of methanol and acetonitrile in the mixture of methanol and acetonitrile includes: 3:1, 1:1, 1:3. The volume ratio of the biological sample protective agent to the fluid sample in the collection tube is 2:1-5:1. The saliva sample containing the biological sample protective agent can be stored at 2-8°C for more than one week to ensure that the biological information of the sample does not change.

[0057] It should be noted that the method of use of the present invention will not affect doctors' use after being protected. It is a preferred operating method and does not fall within the scope of disease treatment methods.

[0058] The following experiments are used to verify the technical effects of the present invention;

[0059] Experiment 1: An experiment to verify the effect of the filter component in the saliva collection device on improving the consistency of the saliva sample;

[0060] 1-1, after adding biological sample protective agent, the saliva sample collected by the device is stored at 2-8°C for 0-7 days, and then centrifuged and the supernatant is diluted 1-100 times for subsequent molecular detection and analysis. The results are as follows Figure 2 .

[0061] When the detection object is a metabolic molecule, the centrifugal force is 2000-10000g, the supernatant diluent is a mixed solvent of methanol, acetonitrile and water, the detection instrument is a mass spectrometer, and the detection molecular weight range is 20-350Da.

[0062] The experimental results are as attached Figure 3 Shown is a representative salivary metabolic profile;

[0063] It can be seen from the figure that the saliva sample obtained by the body fluid collection, pretreatment and storage device of the present invention contains a large amount of metabolic molecule information, which can be obtained by mass spectrometry detection.

[0064] 1-2, using the same individual sample to collect saliva samples three times using a body fluid collection device and a collection device using only a funnel and a collection tube to obtain a saliva metabolic profile and its coefficient of variation (CV) value;

[0065] The experimental results are as attached Figure 4 .

[0066] Result analysis: According to the attached Figure 4It can be seen from the information that the consistency of the saliva sample information obtained using the body fluid collection device is significantly higher than that obtained using the conventional collection device.

[0067] 1-3, metabolic profiles were obtained using saliva samples collected three times from the same individual after passing through a hydrophilic porous polymer filter plate and a hydrophobic porous polymer filter plate;

[0068] The experimental results are as attached Figure 5 As shown;

[0069] Result analysis: It can be seen from the figure that the consistency of the mass spectrometry information of the metabolic samples after passing through the hydrophobic porous polymer filter plate is much better than that of the metabolic samples after passing through the hydrophilic porous polymer filter plate.

[0070] The porous polymer filter plate used in the present invention can intercept and adsorb mucin, cell debris, and insoluble matter in body fluid samples such as saliva, but will not cause the loss of biological information in the body fluid samples.

[0071] 1-4, comparison of metabolic profiles of saliva samples that passed through a microporous membrane without using a porous polymer filter plate;

[0072] The experimental results are as follows Figure 6 As shown;

[0073] Result analysis: Figure 6 It can be seen that there is no significant difference between the two. However, compared with the former, the collection device disclosed in the present invention has less resistance and is simple to operate during the initial purification process of filtration, and is suitable for operation by non-professionals.

[0074] The adsorption and filtration capabilities of the filter component for viscous fluid samples are basically constant. When the volume of the saliva sample entering the sampling tube is 0.5-1 mL, the volume of the clarified saliva sample collected in the collection tube is in the range of 0.15-0.3 mL. Therefore, the body fluid collection device disclosed in the present invention can achieve the collection of saliva samples with a relatively stable volume.

[0075] Experiment 2: an experiment to verify the storage conditions of saliva samples obtained by the device of the present invention;

[0076] The components of human body fluid samples are complex, and blood, urine, saliva, etc. all contain a large amount of proteins, peptides, endogenous metabolites, inorganic ions, and circulating free DNA. Saliva also contains a large amount of bacteria, cell debris, impurities, and oral insoluble matter. The regular storage conditions for biological samples are ultra-low temperature storage, and the laboratory storage temperature is -20°C or -80°C. The saliva collection device of the present invention has a filtering component with a step-by-step filtering function, which can filter out sticky proteins, cell debris, impurities, oral insoluble matter and bacteria at the same time, so the storage conditions can be reduced, which is more suitable for non-centralized sampling.

[0077] To verify this effect, the specific operation steps of this embodiment are as follows:

[0078] Step 1, the subject cleans the oral cavity and calmly produces saliva for 2-5 minutes, and three tubes of saliva samples are continuously collected using the body fluid sample collection device of the present invention;

[0079] Step 2, divide the 3 tubes of saliva samples collected into 3 portions and pack them into equal volumes;

[0080] Step 3, the packaged saliva samples were placed in refrigerators at 2-8°C, -20°C and -80°C for 72 hours;

[0081] Step 4: After 72 hours, the saliva samples stored under different conditions are subjected to metabolic molecule extraction experiments and mass spectrometry analysis, and the spectra of the same sample under different storage conditions are compared and the coefficient of variation is calculated.

[0082] Experimental results: The coefficient of variation of 8 tubes of saliva samples placed under 3 different storage conditions, such as Figure 7 As shown;

[0083] Result analysis: Figure 7 It can be seen that the coefficient of variation of each tube of saliva sample under three different storage conditions was <20%, indicating that the biomolecular information of the saliva samples in a non-frozen state can maintain good consistency with the results under -20℃ and -80℃ storage conditions.

[0084] The body fluid samples collected by the saliva collecting device of the present invention do not need to be stored at low temperatures.

[0085] Experiment 3, verifying the non-cryogenic stability of saliva samples;

[0086] The experimental steps are as follows:

[0087] Step 1, the subject cleans the oral cavity and calmly produces saliva for 2-5 minutes, and three tubes of saliva samples are continuously collected using the body fluid sample collection device of the present invention;

[0088] Step 2, divide the collected 3 tubes of saliva samples into 7 portions and pack them in equal volumes;

[0089] Step 3, adding the biological sample protective agent in the saliva sample collection device of the present invention to the packaged saliva sample, fully shaking and evenly storing at 2-8° C., setting the storage time to 0 hours, 4 hours, 8 hours, 24 hours, 72 hours (3 days), 120 hours (5 days), 168 hours (7 days), and performing a centrifugation step after reaching the set time to volatilize the solvent of the supernatant to form a sample dry powder, which is stored at -20° C. or -80° C.;

[0090] Step 4, the saliva sample dry powder pretreated at different time points is re-dissolved and mass spectrometry analysis is performed, and the spectra of the packaged samples of the same sample placed for 4 hours, 8 hours, 24 hours, 72 hours (3 days), 120 hours (5 days), and 168 hours (7 days) are compared with the spectrum of the sample placed for 0 hours and immediately processed, and the coefficient of variation is calculated.

[0091] Experimental results: Representative spectra of saliva samples placed at different times are shown below: Figure 8 shown.

[0092] Result analysis: Figure 8 It can be seen that within 7 days, the coefficient of variation of the saliva samples was <25%, indicating that the saliva samples collected, pretreated and stored by the saliva collection device of the present invention can maintain stability for a long time under conventional household storage conditions.

[0093] The device adopts a porous polymer filter plate and a filter head filter component, which can not only ensure the collection of non-irritating saliva samples, but also eliminate the interference of mucin, bacteria, cell debris, oral insoluble matter / impurities at one time under low resistance, complete the initial purification process, and improve the stability of biological molecules in the saliva sample, especially metabolic molecules. In particular, the hydrophobic porous polymer filter plate is particularly effective in combination with a low-adsorption microporous filter membrane. Such an effect cannot be achieved by using only a hydrophobic porous polymer filter plate or only a low-adsorption microporous filter membrane. The present invention has innovatively discovered that the combination of the two can achieve such an unexpected effect.

[0094] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the above embodiments do not limit the present invention in any form, and any technical solution obtained by equivalent replacement or equivalent transformation falls within the protection scope of the present invention.

Claims

1. A body fluid collection, pretreatment and storage device, characterized in that: include: A sampling tube, a push tube sliding in the sampling tube, a filter assembly arranged at the bottom of the sampling tube, a collection tube connected to the filter assembly, and a biological sample protective agent; The filter assembly comprises: a porous polymer filter plate arranged inside the sampling tube, and a filter head connected to the lower end of the sampling tube; The filter head comprises: a filter head body, and a low-adsorption microporous filter membrane disposed in the filter head body; the pore size of the low-adsorption microporous filter membrane is 0.2-0.5 μm; The porous polymer filter plate is a hydrophobic porous polymer filter plate, the plane size of the hydrophobic porous polymer filter plate is the same as the inner diameter of the sampling tube, the thickness is 1-3 mm, and the pore size is 10-100 μm.

2. A body fluid collection, pretreatment and storage device according to claim 1, characterized in that: The collecting tube comprises: a collecting tube body, a pointed bottom formed at the bottom of the collecting tube body, a tube cover connected to the collecting tube body, and scales formed on the outer wall of the collecting tube body.

3. A body fluid collection, pretreatment and storage device according to claim 2, characterized in that: The collecting tube body is connected to the sampling tube through threads.

4. A body fluid collection, pretreatment and storage device according to claim 2, characterized in that: The tube opening of the collecting tube body is larger than the diameter of the liquid outlet of the filter head.

5. A body fluid collection, pretreatment and storage device according to claim 1, characterized in that: Also includes: A sampling funnel movably connected to the sampling tube.

6. A body fluid collection, pretreatment and storage device according to claim 5, characterized in that: The connection modes between the sampling tube and the sampling funnel include: threaded connection and glued connection.

7. A body fluid collection, pretreatment and storage device according to claim 1, characterized in that: The biological sample protective agent includes: methanol, ethanol, acetonitrile, acetone, and a mixture of methanol and acetonitrile.

8. A body fluid collection, pretreatment and storage device according to claim 1, characterized in that: The volume ratio of the biological sample protective agent to the sample is 2:1-5:

1.

9. A method for using a body fluid collection, pretreatment and storage device according to claim 1, characterized in that: Step 1: The subject holds the upper end of the body fluid sampling tube and collects the body fluid into the sampling funnel. The body fluid is retained on the porous polymer filter plate. The sampling volume is 0.5-1 mL. Step 2: Separate the sampling funnel from the sampling tube, hold the tail of the push tube, and push the head of the push tube directly into the sampling tube. The sample passes through the porous polymer filter plate and the filter head and enters the collection tube. Step 3, separating the collection tube from the sampling tube, thus obtaining the pre-treated body fluid sample with a volume of 0.15-0.3 mL; Step 4: Pour the biological sample preservative into the collection tube, cover the tube tightly, and mix the biological sample preservative and body fluid sample evenly.

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