CN111705105A - Qualitative detection capability verification sample for hemolytic streptococcus in cotton swab and preparation method thereof - Google Patents
Qualitative detection capability verification sample for hemolytic streptococcus in cotton swab and preparation method thereof Download PDFInfo
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- 238000012795 verification Methods 0.000 title claims abstract description 30
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- 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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/14—Streptococcus; Staphylococcus
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- C12N1/20—Bacteria; Culture media therefor
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/315—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Streptococcus (G), e.g. Enterococci
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Abstract
The invention discloses a qualitative detection capability verification sample of hemolytic streptococcus in a cotton swab and a preparation method thereof. The sample can be used for evaluating the laboratory detection capability, controlling the quality, checking personnel, verifying the method and the like in the detection process, and fills the blank at home and abroad. Meanwhile, the uniformity and the stability of the sample meet the capability verification requirements, and the problem that the viable pathogenic bacteria in the hemolytic streptococcus detection sample in the cotton swab can change in the processes of transportation, storage, testing and the like is solved. In addition, the preparation method provided by the invention has simple process and high success rate of preparing samples meeting the requirements.
Description
Technical Field
The invention relates to the technical field of microbiological inspection quality control, in particular to a qualitative detection capability verification sample of hemolytic streptococcus in a cotton swab and a preparation method thereof.
Background
The cotton swab belongs to a disposable sanitary product, which is various daily living goods which are discarded after being used once, are in direct or indirect contact with a human body and are used for achieving the purposes of physiological sanitation or health care (antibiosis or bacteriostasis) of the human body. The disposable sanitary product directly contacts skin or mucous membrane, and the quality of the disposable sanitary product directly influences the body health of a user.
The capability of qualitatively detecting the microorganisms in the disposable sanitary product not only can directly reflect the degree of the microorganism pollution, but also is a necessary means for evaluating raw materials, tool equipment, process flows and the sanitary condition of operators used in the production process, and is a comprehensive basis for sanitary evaluation of the disposable sanitary product.
Due to the particularity of the field of microbiological examination of disposable sanitary products, no organization/organization has yet prepared a sample for qualitative detection of hemolytic streptococcus using a cotton swab as a matrix at home and abroad, and no related literature reports exist.
Hemolytic streptococcus (β -hemolytic streptococcus) is widely distributed in nature, exists in water, air, dust, feces, and oral cavity, nasal cavity, and throat of healthy people and animals, and can be infected by direct contact, air spray transmission, or wound through skin and mucosa, and contaminated food such as milk, meat, egg, and their products can also infect human. Hemolytic streptococcus often causes pyogenic inflammation of skin, subcutaneous tissue, respiratory tract infections, fulminant epidemic of epidemic pharyngitis and allergic reactions such as neonatal sepsis, bacterial endocarditis, scarlet fever and rheumatic fever, glomerulonephritis. Therefore, a sample for verifying the qualitative detection capability of hemolytic streptococcus in the mask is urgently needed.
Disclosure of Invention
Aiming at the technical problems, the invention provides a qualitative detection capability verification sample of hemolytic streptococcus in a cotton swab and a preparation method thereof, and provides important guarantee for microbiological capability verification of disposable hygienic products.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention firstly provides a qualitative detection capability verification sample of hemolytic streptococcus in a cotton swab, wherein the sample comprises a negative sample and a positive sample, the positive sample contains a matrix, target bacteria and interference bacteria, and the negative sample contains a matrix and interference bacteria; the matrix is cotton swabs, the target bacteria are beta-hemolytic streptococcus (beta-hemolytic streptococcus), and the interfering bacteria are one or more of Listeria lnonensis (Listeria monocytogenes), Enterococcus faecalis (Enterococcus faecalis), Escherichia coli (Escherichia coli), Staphylococcus capitis (Staphylococcus capitis), and Bacillus mycoides (Bacillus mycoides).
In the sample verified by the qualitative detection capability of the hemolytic streptococcus in the cotton swab, the concentration of bacteria in the sample is 103-106cfu/mL。
In the qualitative detection capability verification sample of the hemolytic streptococcus in the cotton swab, the target bacteria and the interference bacteria of the positive sample are mixed into mixed bacteria according to the volume ratio of 1:2-3, and the mixed bacteria are added into the matrix.
In the qualitative detection capability verification sample for the hemolytic streptococcus in the cotton swab, the sample is a freeze-dried sample, and the freeze-drying protective agent contains 8-10% of trehalose, 3-5% of sodium glutamate, 0.5-3% of gelatin and 2-5% of bovine serum albumin.
In the qualitative detection capability verification sample for the hemolytic streptococcus in the cotton swab, the volume ratio of each strain to the freeze-drying protective agent is 1: 9-3: 7.
In the qualitative detection capability verification sample for the hemolytic streptococcus in the cotton swab, the cotton swab is made of absorbent cotton, and the cotton swab is sterilized for later use.
The invention also provides a preparation method of the qualitative detection capability verification sample of the hemolytic streptococcus in the cotton swab, which comprises the following steps:
s1 strain recovery passage
Recovering each strain, and identifying the recovered strain;
s2, Strain amplification
Culturing the recovered single viable strain at 36 + -1 deg.C for 36h, eluting with sterile normal saline, and gradually diluting to 10% concentration3-106cfu/mL, and mixing with a freeze-drying protective agent according to the proportion of 1: 9-3: 7 to prepare freeze-dried bacterial suspension of corresponding single strain;
s3, preparing bacterial suspension
Positive sample according to target concentration of target bacteria 102-105cfu/mL and target concentration of each of the interfering bacteria 102-105Preparing bacterial suspension by cfu/mL, mixing the bacterial suspension and the interfering bacteria according to the volume ratio of 1:2-3 of the target bacteria to the interfering bacteria to prepare mixed bacterial suspension, and continuously stirring the mixed bacterial suspension on a magnetic stirrer;
negative sample interfering bacteria at a target concentration of 10 per bacteria3-106cfu/mL are mixed in equal volume to prepare a mixed bacterial suspension, and the mixed bacterial suspension is placed on a magnetic stirrer to be continuously stirred;
s4 matrix treatment
Taking a disposable cotton swab, wherein the cotton swab is made of absorbent cotton and is used for standby after sterilization;
s5, freeze-drying
And (3) dropwise adding the mixed bacterial suspension into each part of cotton swab matrix, filling the cotton swab matrix into a sample bottle after the cotton swab completely absorbs the liquid, and freeze-drying to obtain a freeze-dried sample.
In the preparation method of the qualitative detection capability verification sample of the hemolytic streptococcus in the cotton swab, in the step S1, each strain is respectively inoculated to a Columbia blood agar culture medium, the target bacteria are subjected to anaerobic culture for 36 hours at the temperature of 36 +/-1 ℃, and the interfering bacteria are subjected to aerobic culture for 36 hours at the temperature of 36 +/-1 ℃.
In the preparation method of the qualitative detection capability verification sample of the hemolytic streptococcus in the cotton swab, the freeze-drying protective agent in the step S2 contains 8-10% of trehalose by volume fraction, 3-5% of sodium glutamate by volume fraction, 0.5-3% of gelatin by volume fraction and 2-5% of bovine serum albumin by volume fraction.
According to the preparation method of the qualitative detection capability verification sample of the hemolytic streptococcus in the cotton swab, the freeze-drying time in the step S5 is 40-50 h.
Compared with the prior art, the invention has the beneficial effects that:
the qualitative detection capability verification sample for the hemolytic streptococcus in the cotton swab is prepared by taking the cotton swab as a matrix and adding target bacteria and/or interference bacteria to prepare a positive sample and a negative sample for the first time, and is different from a capability verification simulation sample in the field of microorganisms with freeze-dried powder properties. The sample can be used for evaluating the laboratory detection capability, controlling the quality, checking personnel, verifying the method and the like in the detection process, and fills the blank at home and abroad. Meanwhile, the uniformity and the stability of the sample meet the capability verification requirements, and the problem that the viable pathogenic bacteria in the hemolytic streptococcus detection sample in the cotton swab can change in the processes of transportation, storage, testing and the like is solved. In addition, the preparation method provided by the invention has simple process, and the success rate of obtaining the sample required by CNAS-GL 03 & lt & gt evaluation guidelines for uniformity and stability of capability verification sample & gt is high.
Detailed Description
The invention provides a qualitative detection capability verification sample of hemolytic streptococcus in a cotton swab, which comprises a negative sample and a positive sample, wherein the positive sample contains a matrix, target bacteria and interference bacteria, and the negative sample contains the matrix and the interference bacteria; the target bacteria are main detection bacteria of the examination sample, and the interference bacteria are bacteria with similar biochemistry or similar shape of the target bacteria. Can accurately separate and distinguish target bacteria and background bacteria as main examination targets.
Wherein the substrate is cotton swabs, the target bacteria is beta hemolytic streptococcus, and the interfering bacteria is one or more of Listeria lnokhei, enterococcus faecalis, Escherichia coli, Staphylococcus capitis and Bacillus mycoides. The beta hemolytic streptococcus (CMCC 32210) adopted by the invention is purchased from CMCC (China medical bacteria collection management center), and Listeria lnonix (ATCC 33090), enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 25922), staphylococcus capitis (ATCC35661) and bacillus mycoides (ATCC 10206) are purchased from ATCC (American type culture Collection) and CMCC (China medical bacteria collection management center), so that the traceability of the strain is ensured, and the strain is only taken as an exemplary example of the implementation mode of the invention and can be replaced by an equivalent strain.
In order to make those skilled in the art better understand the technical solution of the present invention, the present invention will be further described in detail with reference to the following embodiments.
Example 1 preparation of qualitative assay for Streptococcus hemolyticus in Cotton swab
The method comprises the following steps:
s1 strain recovery passage
Respectively inoculating each strain to a Columbia blood agar culture medium, carrying out anaerobic culture at the temperature of 36 +/-1 ℃ for 36h on target bacteria at the temperature of 36 +/-1 ℃, carrying out aerobic culture at the temperature of 36 +/-1 ℃ for 36h on interfering bacteria, and identifying the recovered strains;
s2, Strain amplification
Culturing the recovered single viable strain at 36 +/-1 ℃ for 36h, culturing the target hemolytic streptococcus to a stationary phase, culturing other interfering bacteria to the end of a logarithmic growth phase, wherein the bacteria in the stationary phase and the last logarithmic growth phase have biochemical characteristics more typical than those in the logarithmic growth phase, have stronger resistance to mechanical damage in the freeze drying process, are eluted by using sterile physiological saline, and are gradually diluted until the bacteria concentration is 103cfu/mL, mixing with lyophilized protectant at a ratio of 1:9, and making into single strainFreeze-drying the bacterial suspension; the freeze-drying protective agent contains trehalose 8% by volume, sodium glutamate 3% by volume, gelatin 0.5% by volume, and bovine serum albumin 2% by volume, and is sterilized with sterile water, such as sterile needle filter with pore diameter of 0.22 μm, and used after 2 times of filtration sterilization;
s3, preparing bacterial suspension
Positive sample according to target concentration of target bacteria 102cfu/mL and target concentration of each of the interfering bacteria 102Preparing bacterial suspension by cfu/mL, mixing the bacterial suspension and the interfering bacteria according to the volume ratio of 1:2 of the target bacteria to the interfering bacteria to prepare mixed bacterial suspension, and continuously stirring the mixed bacterial suspension on a magnetic stirrer;
negative sample interfering bacteria at a target concentration of 10 per bacteria3cfu/mL are mixed in equal volume to prepare a mixed bacterial suspension, and the mixed bacterial suspension is placed on a magnetic stirrer to be continuously stirred;
s4 matrix treatment
Taking a disposable cotton swab, wherein the cotton swab is made of absorbent cotton and is used for standby after sterilization;
s5, freeze-drying
The mixed bacteria suspension is dripped into each cotton swab matrix, after the cotton swabs completely absorb liquid, the cotton swabs are filled into sample bottles (penicillin bottles), the sample bottles are placed into a freeze dryer with a cover, freeze drying is carried out for 40 hours, after sample freeze drying is finished, in-box plugging is directly carried out, the machine is closed, samples in the sample bottles are in a vacuum state, and after the machine is shut down, the disposable aluminum covers capable of being torn are pressed outside rubber plugs, so that freeze-dried samples are obtained. The freeze dryer parameters were set as follows: freezing speed is 1 ℃/min; early freezing point-1 deg.C for 15 min; the freezing temperature is-45 ℃; eutectic point-31 ℃; the heating temperature is 15 ℃ for 180 min. Starting the freeze dryer, and directly performing programmed freeze drying process by the machine for 40 h.
S6, packaging and storing
The lyophilized samples were stored at a temperature of 4 ℃.
Example 2 preparation of qualitative assay for Streptococcus hemolyticus in Cotton swab
The method comprises the following steps:
s1 strain recovery passage
Respectively inoculating each strain to a Columbia blood agar culture medium, carrying out anaerobic culture on target bacteria at the temperature of 36 +/-1 ℃ for 36h, carrying out aerobic culture on interference bacteria at the temperature of 36 +/-1 ℃ for 36h, and identifying the recovered strains;
s2, Strain amplification
Culturing the recovered single viable strain at 36 + -1 deg.C for 36h, eluting with sterile normal saline, and gradually diluting to bacteria concentration of 104cfu/mL, mixing with a freeze-drying protective agent according to the proportion of 2:8 to prepare freeze-dried bacterial suspension of corresponding single strain; the freeze-drying protective agent contains trehalose with the volume fraction of 9%, sodium glutamate with the volume fraction of 4%, gelatin with the volume fraction of 1% and bovine serum albumin with the volume fraction of 3%, and is prepared by sterile water for sterilization and then used;
s3, preparing bacterial suspension
Positive sample according to target concentration of target bacteria 103cfu/mL and target concentration of each of the interfering bacteria 103Preparing bacterial suspension by cfu/mL, mixing the bacterial suspension and the interfering bacteria according to the volume ratio of 1:3 of the target bacteria to the interfering bacteria to prepare mixed bacterial suspension, and continuously stirring the mixed bacterial suspension on a magnetic stirrer;
negative sample interfering bacteria at a target concentration of 10 per bacteria4cfu/mL are mixed in equal volume to prepare a mixed bacterial suspension, and the mixed bacterial suspension is placed on a magnetic stirrer to be continuously stirred;
s4 matrix treatment
Taking a disposable cotton swab, wherein the cotton swab is made of absorbent cotton and is used for standby after sterilization;
s5, freeze-drying
The mixed bacteria suspension is dripped into each cotton swab matrix, after the cotton swabs completely absorb liquid, the liquid is filled into a sample bottle (penicillin bottle), the sample bottle is covered and placed into a freeze dryer, the freeze dryer parameters are as in embodiment 1, freeze drying is carried out for 45 hours, after the sample freeze drying is finished, in-box plugging is directly carried out, the machine is closed, the sample in the sample bottle is in a vacuum state, and the aluminum cover can be torn off at one time by pressing the rubber plug outside the machine, so that a freeze-dried sample is obtained.
S6, packaging and storing
The lyophilized samples were stored at a temperature of 4 ℃.
Example 3 preparation of qualitative assay for Streptococcus hemolyticus in Cotton swab
The method comprises the following steps:
s1 strain recovery passage
Respectively inoculating each strain to a Columbia blood agar culture medium, carrying out anaerobic culture on target bacteria at the temperature of 36 +/-1 ℃ for 36h, carrying out aerobic culture on interference bacteria at the temperature of 36 +/-1 ℃ for 36h, and identifying the recovered strains;
s2, Strain amplification
Culturing the recovered single viable strain at 36 + -1 deg.C for 36h, eluting with sterile normal saline, and gradually diluting to bacteria concentration of 106cfu/mL, mixing with a freeze-drying protective agent according to the proportion of 3:7 to prepare freeze-dried bacterial suspension of corresponding single strain; the freeze-drying protective agent contains 10% of trehalose, 5% of sodium glutamate, 3% of gelatin and 5% of bovine serum albumin by volume fraction, and is prepared by sterile water for sterilization;
s3, preparing bacterial suspension
Positive sample according to target concentration of target bacteria 105cfu/mL and target concentration of each of the interfering bacteria 105Preparing bacterial suspension by cfu/mL, mixing the bacterial suspension and the interfering bacteria according to the volume ratio of 1:3 of the target bacteria to the interfering bacteria to prepare mixed bacterial suspension, and continuously stirring the mixed bacterial suspension on a magnetic stirrer;
negative sample interfering bacteria at a target concentration of 10 per bacteria6cfu/mL are mixed in equal volume to prepare a mixed bacterial suspension, and the mixed bacterial suspension is placed on a magnetic stirrer to be continuously stirred;
s4 matrix treatment
Taking a disposable cotton swab, wherein the cotton swab is made of absorbent cotton and is used for standby after sterilization;
s5, freeze-drying
And (2) dropwise adding the mixed bacterial suspension into each part of cotton swab matrix, filling the cotton swab into a sample bottle (penicillin bottle) after the liquid is completely absorbed by the cotton swab, covering the cotton swab in the sample bottle, and putting the cotton swab into a freeze dryer, wherein the freeze dryer has the parameters shown in the embodiment 1, freeze-drying for 50 hours, directly plugging the sample in a box after the sample is freeze-dried, closing the freeze dryer, and keeping the sample in the sample bottle in a vacuum state to obtain a freeze-dried sample.
S6, packaging and storing
The lyophilized samples were stored at a temperature of 4 ℃.
Example 4 demonstration of the homogeneity and stability of Streptococcus hemolyticus in Cotton swab
Uniformity and stability checks of target bacteria in a sample are the main methods to verify the effectiveness of the sample preparation process. Examination of sample uniformity and stability was performed according to CNAS-GL 03 "guidelines for evaluation of uniformity and stability of samples for capability verification".
The uniformity and stability detection method of the obtained sample and the result are as follows:
10 samples of example 1 were randomly selected, spread and inoculated on a Columbia CAN blood agar plate, anaerobically cultured, the target bacteria were counted, and 2X 10 samples of Escherichia coli were tested under repeated conditions. The data of the results were statistically processed by ANOVA, and the statistical procedure and results are shown in Table 1.
TABLE 1 detection of beta hemolytic streptococci in the samples
TABLE 2 statistical results of analysis of variance of beta hemolytic streptococci in samples
And (4) conclusion: critical value of F0.05(9,10)3.02. The calculated F value was 1.78 and the sample F value < the critical value, indicating that at a significant level of 0.05, the distribution of the target bacteria in the sample was uniform. The homogeneity results of the resulting samples of examples 2 and 3 are similar to example 1.
Stability two types of tests were used: one is the stability test at storage temperature (4 ℃) and the other is the stability test at elevated temperature (simulating the transport conditions of the sample), three temperature points were chosen, 25 ℃, 36 ℃ and 42 ℃. The sample of the embodiment 2 is randomly taken, the sample is periodically detected, 3 samples are tested according to different storage time of different temperature points, and when the detection value of the sample in the stability test is consistent with the specified value, the stability of the capability verification sample is in accordance with the requirement, and meanwhile, the longest storage time in accordance with the requirement of the capability verification sample under different temperature conditions is determined. The stability test results are shown in table 3 below.
TABLE 3 stability test results
And (4) conclusion: the sample is stored at the low temperature of 4 ℃ for 120 days and at the high temperature of 42 ℃ for 20 days, the detection results of the sample all meet the requirements, and the stability is good.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: it is to be understood that modifications may be made to the technical solutions described in the foregoing embodiments, or equivalents may be substituted for some of the technical features thereof, but such modifications or substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. The qualitative detection capability verification sample of the hemolytic streptococcus in the cotton swab is characterized by comprising a negative sample and a positive sample, wherein the positive sample contains a matrix, target bacteria and interference bacteria, and the negative sample contains the matrix and the interference bacteria; the matrix is cotton swabs, the target bacteria is beta hemolytic streptococcus, and the interference bacteria is one or more of Listeria lnokhei, enterococcus faecalis, Escherichia coli, staphylococcus capitis and Bacillus mycoides.
2. The qualitative test for the ability to detect hemolytic streptococci in a cotton swab of claim 1, wherein the concentration of bacteria in the sample is 103-106cfu/mL。
3. The qualitative detection capability verification sample of hemolytic streptococcus in cotton swab of claim 1, wherein the target bacteria and the interfering bacteria of the positive sample are mixed into mixed bacteria according to a volume ratio of 1:2-3, and the mixed bacteria are added into the matrix.
4. The qualitative test capability verification sample of hemolytic streptococcus in cotton swab of claim 1, wherein the sample is a freeze-dried sample, and the freeze-drying protective agent comprises trehalose with a volume fraction of 8-10%, sodium glutamate with a volume fraction of 3-5%, gelatin with a volume fraction of 0.5-3%, and bovine serum albumin with a volume fraction of 2-5%.
5. The qualitative detection capability verification sample of hemolytic streptococcus in cotton swab of claim 4, wherein the volume ratio of each strain to the freeze-drying protective agent is 1: 9-3: 7.
6. The qualitative test capability verification sample of hemolytic streptococcus in cotton swab of claim 1, wherein the cotton swab is made of absorbent cotton, and the cotton swab is sterilized for later use.
7. The method for preparing the qualitative detection capability verification sample of hemolytic streptococcus in cotton swab of claim 1, comprising the following steps:
s1 strain recovery passage
Recovering each strain, and identifying the recovered strain;
s2, Strain amplification
Culturing the recovered single viable strain at 36 + -1 deg.C for 36h, eluting with sterile normal saline, and gradually diluting to bacteria concentration of 103-106cfu/mL, and mixing with a freeze-drying protective agent according to the proportion of 1: 9-3: 7 to prepare freeze-dried bacterial suspension of corresponding single strain;
s3, preparing bacterial suspension
Positive sample according to target concentration of target bacteria 102-105cfu/mL and target concentration of each of the interfering bacteria 102-105cfu/mL ofPreparing bacterial suspension, mixing the target bacteria and the interfering bacteria according to the volume ratio of 1:2-3 to prepare mixed bacterial suspension, and continuously stirring the mixed bacterial suspension on a magnetic stirrer;
negative sample interfering bacteria at a target concentration of 10 per bacteria3-106cfu/mL are mixed in equal volume to prepare a mixed bacterial suspension, and the mixed bacterial suspension is placed on a magnetic stirrer to be continuously stirred;
s4 matrix treatment
Taking a disposable cotton swab, wherein the cotton swab is made of absorbent cotton and is used for standby after sterilization;
s5, freeze-drying
And (3) dropwise adding the mixed bacterial suspension into each part of cotton swab matrix, filling the cotton swab matrix into a sample bottle after the cotton swab completely absorbs the liquid, and freeze-drying to obtain a freeze-dried sample.
8. The method for preparing a sample for verifying the qualitative detection capability of hemolytic streptococcus in a cotton swab of claim 1, wherein in step S1, each strain is inoculated to a Columbia blood agar culture medium, the target bacteria are anaerobically cultured at a temperature of 36 ℃ ± 1 ℃ for 36 hours, and the interfering bacteria are aerobically cultured at a temperature of 36 ℃ ± 1 ℃ for 36 hours.
9. The method for preparing a sample for verifying the qualitative detection capability of hemolytic streptococcus in a cotton swab of claim 1, wherein the lyoprotectant in step S2 comprises trehalose in an amount of 8-10% by volume, sodium glutamate in an amount of 3-5% by volume, gelatin in an amount of 0.5-3% by volume, and bovine serum albumin in an amount of 2-5% by volume.
10. The method for preparing a sample for verifying the qualitative detection capability of Streptococcus hemolyticus in a cotton swab according to claim 1, wherein the lyophilization time in step S5 is 40-50 hours.
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