Formula of wetting liquid for improving sample preservation effect
By adding anti-aggregation reagents, buffers and other ingredients to the wetting solution, the problem of cell aggregation and drying during sample collection and preservation is solved, and efficient storage and quality assurance of samples is achieved.
Patent Information
- Application Number
- CN202411870879.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-18
- Publication Date
- 2025-03-14
AI Technical Summary
During the sample collection and storage process, existing wetting fluids can easily lead to cell aggregation, sedimentation or drying and inactivation, affecting virus activity and sample quality.
A wetting solution formula containing anti-aggregation reagents, buffers, ionic strength maintenance reagents, antimicrobial reagents, mucus dissolution reagents, detergents and penicillanes is provided. Through the combination of these ingredients, cells are prevented from aggregation and drying and maintained the activity and quality of the sample.
Effectively prevent the aggregation, sedimentation or drying and inactivation of the shedded cells, maintain the integrity of the cell morphology, improve the survival rate of cells, ensure the normal progress of the enzymatic reaction, control the osmotic pressure, inhibit the growth of microorganisms, decompose mucus, remove impurities, and prolong the effective storage time of the sample.
Smart Images
Figure CN119614675A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of medical testing, in particular to a wetting liquid formula for increasing sample preservation effect. Background Art
[0002] HPV testing for men is of great significance in controlling the incidence of cervical cancer. However, HPV testing for men in hospitals uses urethral collection or prostatic fluid as sample sources, which is very difficult and painful. Later, researchers found that collecting samples from the coronal sulcus of the male penis is more efficient because there are more shed cells in this part and they are easy to aggregate, and HPV samples in the smegma are more concentrated. However, if a dry brush is used to collect samples directly, it is difficult to adhere and it is very painful.
[0003] Since the collection process may cause drying or aggregation problems, which will affect the activity of the virus, the current preservation method on the market is mainly to add wetting fluid to ensure the activity of the sample.
[0004] However, the wetting fluids currently available on the market still have a significant impact on the sample transfer and preservation process, such as nucleases that weaken viruses and destroy structures. Therefore, the exploration of wetting fluid preparation methods is still ongoing. Summary of the invention
[0005] In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide a wetting liquid formula that increases the sample preservation effect.
[0006] The technical solution of the present invention is as follows:
[0007] A wetting solution formula for increasing the sample preservation effect, the formula comprising: 15%-50% anti-aggregation agent; 5%-10% buffer; 1%-20% ionic strength maintenance agent; 0.01%-0.5% antimicrobial agent; 0.1%-5% mucolytic agent; 0%-0.5% detergent, 0.1%-0.5% penicillin; the content percentages are calculated based on the amount of 1L solution.
[0008] Furthermore, the anti-aggregation agent mainly functions to prevent the exfoliated cells in the agent from aggregating and settling or drying and inactivating, and mainly includes one or more of glycerol, glycerol monooleate, PEG6000, sorbitol, and propylene glycol; preferably, the concentration of glycerol monooleate is 0.5%-5%; the concentration of PEG 6000 is 0.5%-5%.
[0009] Furthermore, the main function of the buffer is to adjust the pH of the reagent and provide the required ions for the enzymatic reaction, mainly including disodium hydrogen phosphate, sodium acetate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium bicarbonate, sodium hydroxide, etc.; preferably disodium hydrogen phosphate, with a concentration of 1%-10%; preferably sodium hydroxide, with a concentration of 0.01%-1%.
[0010] Furthermore, the ionic strength maintaining agent mainly refers to adding sodium chloride salt solution or amino acid solution to control the osmotic pressure of the overall solution to avoid cell inactivation; the concentration of the sodium chloride solution is 0.01%-10%; the concentration of the amino acid solution is 0.01%-10%, wherein the amino acids include histidine, lysine, arginine, asparagine, aspartic acid, threonine, proline, cytosine nucleoside,
[0011] Cysteine, cytosine, prolylarginine, glutamine, glutamic acid, glutamine, etc.; preferably, the concentration of the amino acid solution is 0.01%-5%, preferably histidine and / or arginine.
[0012] Furthermore, the antimicrobial agent mainly includes EDTA, PCR antifreeze, guanidine thiocyanate, sodium acetate, sodium thiocyanate, hydrochloric acid, etc., preferably, the concentration of EDTA is 0.01%-0.5%; the concentration of PCR antifreeze is 0.01%-0.5%; the concentration of guanidine thiocyanate is 0.01%-0.5%.
[0013] Furthermore, the mucolytic reagent mainly includes trypsin, collagenase, nuclease, etc.; preferably, the concentration of trypsin is 0.1%-1%; preferably, the concentration of collagenase is 0.2%-30%; preferably, the concentration of nuclease is 0.1%-0.5%;
[0014] The cleaning agent mainly includes a surfactant, a dodecyl quaternary ammonium salt, and preferably the concentration of hemoglobin chloride is 0.02%-0.5%;
[0015] The pH value of the wetting liquid is 2.5-3.5; the conductivity is 20-120Sm, and the osmotic pressure is 200-2000mOsm;
[0016] The penicillin is ascorbic acid, preferably at a concentration of 0.1%-1%.
[0017] Furthermore, the order of adding glycerol monooleate and PEG6000 is to add glycerol monooleate first, mix well, and then add PEG6000.
[0018] Furthermore, the order of adding trypsin, collagenase, nuclease, EDTA and penicillin is to add trypsin first, mix well, and then add collagenase, nuclease, penicillin solution and EDTA solution.
[0019] A method for preparing a wetting liquid preparation for increasing sample preservation effect according to any one of claims 1 to 8, characterized in that it comprises the following steps:
[0020] 1) preparing the mother solution of the cleaning and wetting solution;
[0021] 2) preparing a wetting solution preparation;
[0022] 2) Add the sample collection stick and seal.
[0023] Furthermore, in the step 1, a mother solution of the cleaning and wetting solution is prepared, wherein the mother solution comprises an anti-aggregation agent, a buffer, an ionic strength maintaining agent, an antimicrobial agent, a mucolytic agent, a detergent, and penicillin;
[0024] The method for preparing the mother solution is as follows: a. adding the anti-aggregation reagent to the reaction bottle, stirring for 2-5 minutes, adding the buffer solution, stirring for 1-5 minutes; adding the ionic strength maintaining reagent;
[0025] b. Stir for 1-5 minutes, add the mucolytic agent, stir for 5-10 minutes, add penicillin, stir for 2-5 minutes, add the antimicrobial agent and stir for 1-5 minutes;
[0026] In the step 1, during the reaction of the mother solution a or b, the mother solution components are added to the reaction bottle with a stirring speed of 5-100 r / min;
[0027] In the step 1, the mother liquid is prepared and filtered through a filter, and then pasteurized or sterilized;
[0028] In said step 1, the mother solution is added in the order of the groups when it is prepared; preferably, in said step 2, preparing the wetting solution preparation, the mother solution is filled into a disposable sterile sampling brush, and the volume of the mother solution after filling accounts for 75%-95% of the bottle;
[0029] In step 3, the sample collection stick is sterilized before use.
[0030] Compared with the traditional technology, the present invention has the following advantages including but not limited to:
[0031] 1. By adding anti-aggregation reagents, the detached cells can be effectively prevented from aggregating and settling in the solution or drying and inactivation, thereby maintaining the integrity of the cell morphology and improving the cell survival rate.
[0032] 2. The buffer can maintain the pH value in the wetting liquid within an appropriate range, ensuring the normal progress of the enzymatic reaction and providing a stable living environment for the cells.
[0033] 3. Ionic strength maintenance reagents help control the osmotic pressure of the solution, avoiding cell rupture or dehydration caused by osmotic pressure mismatch, thereby protecting cell structure and function.
[0034] 4. Adding antimicrobial agents can effectively inhibit the growth of bacteria, fungi and other microorganisms, reduce the risk of sample contamination, and ensure sample quality.
[0035] 5. Mucolytic reagents can effectively decompose the mucus components in the sample, making the sample easier to handle and analyze.
[0036] 6. The use of detergents can help remove impurities that may exist on the sample surface and improve sample purity.
[0037] 7. As an antioxidant, penicillin can fight free radicals, reduce the damage caused by oxidative stress to samples, and extend the effective storage time of samples.
[0038] Instruction Manual
[0039] Figure 1 It is a comparison diagram of Examples 1-6 and Comparative Examples 1-2 of the present invention;
[0040] Figure 2 It is a comparison diagram of Example 1 and Comparative Examples 3-6 of the present invention; DETAILED DESCRIPTION
[0041] Here, exemplary embodiments will be described in detail, and unless otherwise indicated, the embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present disclosure. Instead, they are only examples consistent with some aspects of the present disclosure as detailed in the appended claims.
[0042] Embodiment 1:
[0043] Preparation of wetting fluid formulations
[0044] Preparation of wetting solution:
[0045] Glycerol 15%, glycerol monooleate 0.5%, PEG60002%, disodium hydrogen phosphate 10%, sodium hydroxide 0.01%, sodium chloride solution 3%, histidine 5%, EDTA 1%, trypsin 0.5%, collagenase 2%, nuclease 0.1%, hemoglobin chloride 0.02%, antibacterial agent 0.5%, penicillin 0.5%.
[0046] The mother liquor preparation method is as follows: 1) adding 15% glycerol, 0.5% glycerol monooleate, and 2% PEG6000 to a reaction bottle, stirring for 2-5 minutes, adding 10% disodium hydrogen phosphate, and stirring for 1-5 minutes;
[0047] 2) Add 3% sodium chloride and stir for 1-5 minutes, add 5% histidine and stir evenly, add 1% EDTA and stir for 1-5 minutes.
[0048] 3) adding 0.5% antimicrobial agent, 0.5% trypsin, and then adding 1% EDTA in batches, followed by 2% collagenase and 0.1% nuclease;
[0049] 4) Add 0.02% hemoglobin chloride and 0.5% penicillin;
[0050] 5) After the stirring reaction is completed, the mixture is filtered through a filter and then pasteurized;
[0051] Preparation of wetting fluid formulations
[0052] Fill the wetting liquid mother solution into a disposable sterile sampling brush, and the volume of the mother solution after filling accounts for 90% of the bottle.
[0053] Embodiment 2:
[0054] Preparation of wetting fluid formulations
[0055] Preparation of wetting solution:
[0056] Glycerol 15%, glycerol monooleate 0.5%, PEG60002%, disodium hydrogen phosphate 10%, sodium hydroxide 0.01%, sodium chloride solution 3%, histidine 5%, EDTA 1%, trypsin 0.5%, collagenase 2%, nuclease 0.1%, hemoglobin chloride 0.02%, antibacterial agent 0.5%, penicillin 0.5%.
[0057] The mother solution preparation method is as follows: 1) add 15% glycerol, 0.5% glycerol monooleate, and 2% PEG6000 to a reaction bottle and keep stirring for 2-5 minutes;
[0058] 2) Add 10% disodium hydrogen phosphate and stir for 1-5 minutes; add 3% sodium chloride and stir for 1-5 minutes, add 5% histidine and stir evenly, add 1% EDTA and stir for 1-5 minutes.
[0059] 3) adding trypsin 0.5% in batches, followed by EDTA 1%, followed by collagenase 2% and nuclease 0.1% in batches;
[0060] 4) Add 0.02% hemoglobin chloride and 0.5% penicillin;
[0061] 5) After the stirring reaction is completed, the mixture is filtered through a filter and then pasteurized;
[0062] Preparation of wetting fluid formulations
[0063] Fill the wetting liquid stock solution into a disposable sterile sampling brush, and the total liquid volume after filling accounts for 90% of the bottle.
[0064] Embodiment 3:
[0065] Preparation of wetting fluid formulations
[0066] Preparation of wetting solution:
[0067] Glycerol 15%, glycerol monooleate 0.5%, PEG60002%, disodium hydrogen phosphate 10%, sodium hydroxide 0.01%, sodium chloride solution 3%, histidine 5%, EDTA 1%, trypsin 0.5%, collagenase 2%, nuclease 0.1%, hemoglobin chloride 0.02%, antibacterial agent 0.5%, penicillin 0.5%.
[0068] The mother solution preparation method is as follows: 1) add 15% glycerol, 0.5% glycerol monooleate, and 2% PEG6000 to a reaction bottle and keep stirring for 2-5 minutes;
[0069] 2) Add 10% disodium hydrogen phosphate and stir for 1-5 minutes; add 3% sodium chloride and stir for 1-5 minutes, add 5% histidine and stir evenly, add 1% EDTA and stir for 1-5 minutes.
[0070] 3) adding 0.5% trypsin, followed by 2% collagenase and 0.1% nuclease in batches;
[0071] 4) Add 0.02% hemoglobin chloride and 0.5% penicillin;
[0072] 5) After the stirring reaction is completed, the mixture is filtered through a filter and then pasteurized;
[0073] Preparation of wetting fluid formulations
[0074] Fill the wetting solution mother solution into a disposable sterile sampling brush, and the volume of the mother solution after filling is 90%.
[0075] Embodiment 4:
[0076] Preparation of wetting fluid formulations
[0077] Preparation of wetting solution:
[0078] Glycerol 15%, glycerol monooleate 0.5%, PEG60002%, disodium hydrogen phosphate 10%, sodium hydroxide 0.01%, sodium chloride solution 3%, histidine 5%, EDTA 1%, trypsin 0.5%, collagenase 2%, nuclease 0.1%, antibacterial agent 0.5%, penicillin 0.5%.
[0079] The mother solution preparation method is as follows: 1) add 15% glycerol, 0.5% glycerol monooleate, and 2% PEG6000 to a reaction bottle and keep stirring for 2-5 minutes;
[0080] 2) Add 10% disodium hydrogen phosphate and stir for 1-5 minutes; add 3% sodium chloride solution and stir for 1-5 minutes, add 5% histidine and stir evenly, add 1% EDTA and stir for 1-5 minutes.
[0081] 3) adding 0.5% trypsin, followed by 2% collagenase and 0.1% nuclease in batches;
[0082] 4) Add 0.5% of penicillin;
[0083] 5) After the stirring reaction is completed, the mixture is filtered through a filter and then pasteurized;
[0084] Preparation of wetting fluid formulations
[0085] Fill a disposable sterile sampling brush with 95% of the wetting solution stock.
[0086] Embodiment 5:
[0087] Preparation of wetting fluid formulations
[0088] Preparation of wetting solution:
[0089] Glycerol 15%, glycerol monooleate 0.5%, PEG60002%, disodium hydrogen phosphate 10%, sodium hydroxide 0.01%, sodium chloride solution 3%, histidine 5%, EDTA 1%, trypsin 0.5%, collagenase 2%, nuclease 0.1%, antibacterial agent 0.5%, penicillin 0.5%.
[0090] The mother solution preparation method is as follows: 1) add 15% glycerol, 0.5% glycerol monooleate, and 2% PEG6000 to a reaction bottle and keep stirring for 2-5 minutes;
[0091] 2) Add 10% disodium hydrogen phosphate and stir for 1-5 minutes; add 3% sodium chloride solution and stir for 1-5 minutes, add 5% histidine and stir evenly, add 1% EDTA and stir for 1-5 minutes.
[0092] 3) adding 0.5% trypsin, followed by 2% collagenase and 0.1% nuclease in batches;
[0093] 4) Add 0.5% of penicillin;
[0094] 5) After the stirring reaction is completed, the mixture is filtered through a filter and then pasteurized;
[0095] Preparation of wetting fluid formulations
[0096] Fill the wetting solution mother solution into a disposable sterile sampling brush, and the volume of the mother solution after filling accounts for 95% of the bottle.
[0097] Embodiment 6:
[0098] Preparation of wetting fluid formulations
[0099] Preparation of wetting solution:
[0100] Glycerol 15%, glycerol monooleate 0.5%, PEG60002%, disodium hydrogen phosphate 10%, sodium hydroxide 0.01%, sodium chloride solution 3%, histidine 5%, EDTA 1%, trypsin 0.5%, collagenase 2%, nuclease 0.1%, antibacterial agent 0.5%, penicillin 0.5%.
[0101] The mother solution preparation method is as follows: 1) add 15% glycerol, 0.5% glycerol monooleate, and 2% PEG6000 to a reaction bottle and keep stirring for 2-5 minutes;
[0102] 2) Add 10% disodium hydrogen phosphate and stir for 1-5 minutes; add 3% sodium chloride solution and stir for 1-5 minutes, add 5% histidine and stir evenly, add 1% EDTA and stir for 1-5 minutes.
[0103] 3) 0.5% trypsin was added in sequence, followed by 2% collagenase and 0.1% nuclease in batches.
[0104] 4) Add 0.5% of penicillin;
[0105] 5) After the stirring reaction is completed, the mixture is filtered through a filter and then pasteurized;
[0106] Preparation of wetting fluid formulations
[0107] Fill the wetting solution mother solution into a disposable sterile sampling brush, and the volume of the mother solution after filling accounts for 95% of the bottle.
[0108] Comparative Example 1:
[0109] Compared with Example 2, glycerol monooleate was not added, and other conditions were the same.
[0110] Comparative Example 2:
[0111] Compared with Example 4, no sodium hydroxide was added. The other conditions were the same.
[0112] Experimental Example 1
[0113] Taking the free base content in the sample DNA after collection as an indicator (the higher the free base content, the worse the sample preservation effect), the sample preservation effects in Examples 1 to 6 and Comparative Examples 1 to 2 were tested.
[0114] The collected samples of Examples 1 to 6 and Comparative Examples 1 to 5 were frozen under the same conditions, respectively, and the changes in free base contents were compared.
[0115] according to Figure 1 It can be seen that the wetting solution of the present invention greatly slows down the decomposition rate of free bases in DNA over time due to the addition of glycerol monooleate and sodium hydroxide during the freezing process. Comparison of Comparative Example 1 with Example 2 shows that the free base content increases rapidly during the freezing process without the addition of glycerol monooleate, indicating that the present invention has a synergistic effect in the preservative of glycerol monooleate; Comparative Example 2 with Example 4 shows that the absence of the addition of sodium hydroxide has a greater impact on the free base content during the freezing process, indicating that the sodium hydroxide of the present invention plays an important role in maintaining the osmotic pressure of the system and the stability of the base.
[0116] Comparative Example 3:
[0117] (1) Compared with Example 1, no disodium hydrogen phosphate was added, and the other conditions were the same.
[0118] (2) Compared with Example 1, histidine was not added, and the other conditions were the same.
[0119] (3) Compared with Example 1, no sodium chloride was added, and the other conditions were the same.
[0120] (4) Compared with Example 1, EDTA was not added, and the other conditions were the same.
[0121] Experimental Example 2
[0122] By comparing Example 1 with Comparative Examples 3 to 6, the free base content in the sample DNA after collection was used as an indicator to evaluate the sample preservation effect in the Example and the Comparative Examples.
[0123] The collected samples of Example 1 and Comparative Examples 3 to 6 were cryopreserved under the same conditions, and the changes in free base content were compared. Figure 2 shown.
[0124] according to Figure 2 It can be seen that the wetting solution described in this patent, during the freezing process, reduces the decomposition rate of the free base content over time due to the addition of disodium hydrogen phosphate, histidine, sodium chloride and EDTA. In comparative examples 3 to 6, there are some components in the sample DNA that are inactivated quickly, indicating that due to the lack of key components, the free base content increases quickly after freezing, which illustrates the synergistic effect of the components described in the present invention.
[0125] Experimental Example 3
[0126] The wetting liquid prepared in Example 2 of the present invention was tested and verified. The verification results showed that the bacterial survival rate was 0 and no bacterial colonies were generated in the sample.
[0127] Experimental Example 4
[0128] The wetting liquid prepared in Example 3 of the present invention was tested and verified. The verification results showed that the bacterial survival rate was 0 and no bacterial colonies were generated in the sample.
[0129] Experimental Example 5
[0130] The wetting liquid prepared in Example 4 of the present invention was tested and verified. The verification results showed that the bacterial survival rate was 0 and no bacterial colonies were generated in the sample.
[0131] Experimental Example 6
[0132] The wetting liquid prepared in Example 5 of the present invention was tested and verified. The verification results showed that the bacterial survival rate was 0 and no bacterial colonies were generated in the sample.
[0133] The above is only one of the specific implementations of the present invention, but the technical features of the present invention are not limited thereto. Any modification, equivalent replacement or improvement made within the technical concept of the present invention shall be included in the protection scope of the present invention.
Claims
1. A wetting liquid formula for increasing sample preservation effect, characterized in that: The formula comprises: anti-aggregation agent 15%-50%; buffer 5%-10%; ionic strength maintenance agent 1%-20%; antimicrobial agent 0.01%-0.5%; mucolytic agent 0.1%-5%; detergent 0%-0.5%; penicillin 0.1%-0.5%; The content percentage is calculated based on the amount of 1L solution.
2. A wetting liquid formula for increasing sample preservation effect according to claim 1, characterized in that: The anti-aggregation agent mainly functions to prevent the exfoliated cells in the agent from aggregating and settling or drying and inactivating, and mainly comprises one or more of glycerol, glycerol monooleate, PEG6000, sorbitol, and propylene glycol; preferably, the concentration of glycerol monooleate is 0.5%-5%; the concentration of PEG6000 is 0.5%-5%.
3. A wetting liquid formula for increasing sample preservation effect according to claim 1, characterized in that: The main function of the buffer is to adjust the pH of the reagent and provide the required ions for the enzymatic reaction. It mainly includes disodium hydrogen phosphate, sodium acetate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium bicarbonate, sodium hydroxide, etc.; preferably disodium hydrogen phosphate, with a concentration of 1%-10%; preferably sodium hydroxide, with a concentration of 0.01%-1%.
4. A wetting liquid formula for increasing sample preservation effect according to claim 1, characterized in that: The ionic strength maintaining reagent mainly refers to adding sodium chloride salt solution or amino acid solution to control the osmotic pressure of the overall solution to avoid cell inactivation; the concentration of the sodium chloride solution is 0.01%-10%; the concentration of the amino acid solution is 0.01%-10%, wherein the amino acids include histidine, lysine, arginine, asparagine, aspartic acid, threonine, proline, cytosine, cysteine, cytosine, prolylarginine, glutamine, glutamic acid, glutamine, etc.; preferably, the concentration of the amino acid solution is 0.01%-5%, preferably histidine and / or arginine.
5. A wetting liquid formula for increasing sample preservation effect according to claim 1, characterized in that: The antimicrobial agent mainly comprises EDTA, PCR antifreeze, guanidine thiocyanate, sodium acetate, sodium thiocyanate, hydrochloric acid, etc., preferably, the concentration of EDTA is 0.01%-0.5%; the concentration of PCR antifreeze is 0.01%-0.5%; the concentration of guanidine thiocyanate is 0.01%-0.5%.
6. A wetting liquid formula for increasing sample preservation effect according to claim 1, characterized in that: The mucolytic reagent mainly includes trypsin, collagenase, nuclease, etc.; preferably, the concentration of trypsin is 0.1%-1%; preferably, the concentration of collagenase is 0.2%-30%; preferably, the concentration of nuclease is 0.1%-0.5%; The cleaning agent mainly includes a surfactant, a dodecyl quaternary ammonium salt, and preferably the concentration of hemoglobin chloride is 0.02%-0.5%; The pH value of the wetting liquid is 2.5-3.5; the conductivity is 20-120Sm, and the osmotic pressure is 200-2000mOsm; The penicillin is ascorbic acid, preferably at a concentration of 0.1%-1%.
7. A wetting liquid formula for increasing sample preservation effect according to claim 2, characterized in that: The order of adding glycerol monooleate and PEG6000 is to first add glycerol monooleate, mix well, and then add PEG6000.
8. A wetting liquid formula for increasing sample preservation effect according to claim 1, characterized in that: The order of adding the trypsin, collagenase, nuclease, EDTA and penicillin is to add trypsin first, mix well, and then add collagenase, nuclease, penicillin solution and EDTA solution.
9. A method for preparing a wetting liquid preparation for increasing sample preservation effect according to any one of claims 1 to 8, characterized in that: The following steps are involved: 1) preparing the mother solution of the cleaning and wetting solution; 2) preparing a wetting solution preparation; 2) Add the sample collection stick and seal.
10. The method according to claim 9, characterized in that: In the step 1, a mother solution of a cleaning and wetting solution is prepared, wherein the mother solution comprises an anti-aggregation agent, a buffer, an ionic strength maintaining agent, an antimicrobial agent, a mucolytic agent, a detergent, and penicillin; The method for preparing the mother solution is as follows: a. adding the anti-aggregation reagent to the reaction bottle, stirring for 2-5 minutes, adding the buffer solution, stirring for 1-5 minutes; adding the ionic strength maintaining reagent; b. Stir for 1-5 minutes, add the mucolytic agent, stir for 5-10 minutes, add penicillin, stir for 2-5 minutes, add the antimicrobial agent and stir for 1-5 minutes; In the step 1, during the reaction of the mother solution a or b, the mother solution components are added to the reaction bottle with a stirring speed of 5-100 r / min; In the step 1, the mother liquid is prepared and filtered through a filter, and then pasteurized or sterilized; In said step 1, the mother solution is added in the order of the groups when it is prepared; preferably, in said step 2, preparing the wetting solution preparation, the mother solution is filled into a disposable sterile sampling brush, and the volume of the mother solution after filling accounts for 75%-95% of the bottle; In step 3, the sample collection stick is sterilized before use.