CN109971673A - Clostridium sporogenes employment and suitability test (E & ST) bacterial strain and preparation method thereof - Google Patents
Clostridium sporogenes employment and suitability test (E & ST) bacterial strain and preparation method thereof Download PDFInfo
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- 230000001580 bacterial effect Effects 0.000 title claims abstract description 54
- 241000193470 Clostridium sporogenes Species 0.000 title claims abstract description 37
- 238000010971 suitability test Methods 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 241000894006 Bacteria Species 0.000 claims abstract description 57
- 238000000034 method Methods 0.000 claims abstract description 25
- 229940079593 drug Drugs 0.000 claims abstract description 16
- 239000003814 drug Substances 0.000 claims abstract description 16
- 238000001514 detection method Methods 0.000 claims abstract description 9
- 244000005700 microbiome Species 0.000 claims abstract description 6
- 238000004108 freeze drying Methods 0.000 claims description 29
- 238000013112 stability test Methods 0.000 claims description 15
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 11
- 108010010803 Gelatin Proteins 0.000 claims description 11
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 11
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 11
- 229920000159 gelatin Polymers 0.000 claims description 11
- 239000008273 gelatin Substances 0.000 claims description 11
- 235000019322 gelatine Nutrition 0.000 claims description 11
- 235000011852 gelatine desserts Nutrition 0.000 claims description 11
- 238000007689 inspection Methods 0.000 claims description 8
- 238000011084 recovery Methods 0.000 claims description 8
- 239000000725 suspension Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 239000008223 sterile water Substances 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 4
- 241000193403 Clostridium Species 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000003760 magnetic stirring Methods 0.000 claims description 4
- 239000011159 matrix material Substances 0.000 claims description 4
- 238000007619 statistical method Methods 0.000 claims description 4
- 241001411320 Eriogonum inflatum Species 0.000 claims description 3
- 230000009191 jumping Effects 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 229910052757 nitrogen Inorganic materials 0.000 claims description 2
- 238000012360 testing method Methods 0.000 abstract description 24
- 230000005764 inhibitory process Effects 0.000 abstract description 2
- 230000002906 microbiologic effect Effects 0.000 abstract description 2
- 238000003908 quality control method Methods 0.000 abstract description 2
- 239000002609 medium Substances 0.000 description 4
- JGSARLDLIJGVTE-UHFFFAOYSA-N 3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid Chemical compound O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000006641 stabilisation Effects 0.000 description 3
- 238000011105 stabilization Methods 0.000 description 3
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000005496 eutectics Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 241001474374 Blennius Species 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000012009 microbiological test Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000012430 stability testing Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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Abstract
The invention belongs to the field of quality control in terms of drug microbiologic inhibition tests, in particular to a kind of clostridium sporogenes employment and suitability test (E & ST) bacterial strain and preparation method thereof.Clostridium sporogenes employment and suitability test (E & ST) bacterial strain only includes a certain amount of clostridium sporogenes.Uniformity of the present invention, stability meet employment and suitability test (E & ST) bacterial strain requirement, bacterium number contained by bacterial strain utmostly meets the needs such as employment and suitability test (E & ST) during drug microorganism detection, growth promotion test, the preparation method of sample within the scope of employment and suitability test (E & ST) standard requirements, simple process, success rate are high.
Description
Technical field
The invention belongs to the field of quality control in terms of microbiologic inhibition tests, in particular to a kind of clostridium sporogenes applicability examination
Test bacterial strain and preparation method thereof.
Background technique
Between the particularity of drug, drug microbiological Test field it is very special, it is desirable that every time detection before cultivate
The test samples such as base are both needed to carry out applicability inspection, have defined concentration range requirement to test bacterial strain.Current existing pure bacterium
Strain is most of not to be quantified, only for qualitative use, only a small number of quantitative bacterial strains.Bacterial strain, uniformity are quantified for such
It is particularly important with stability, the problems such as that there are stability is poor for existing quantitative bacterial strain, and traffic condition is more demanding.
Clostridium sporogenes can generate spore, and Gram-positive moves by periflagellum, and be a kind of shuttle of strictly anaerobic
Bacterium.If in the clostridium sporogenes sample of preparation, clostridium sporogenes easily changes (breeding and death etc.), the guarantor of sample will lead to
Deposit that the phase is short, the uniformity and stability of sample are poor.Therefore, comprehensively consider the biochemical characteristic of bacterium, it is metastable to choose property
Clostridium sporogenes is particularly significant to the uniformity and stability that guarantee sample.The process specifications of clostridium sporogenes sample preparation compare
Height, and the uniformity of sample and stability and the strain of freeze-drying, the process conditions of freeze-drying, the use of freeze drying protectant, again water
The medium etc. of change has close relationship.
Clostridium sporogenes index in drug directly affects the hygienic quality and safety of drug.Therefore, uniformity and stabilization are prepared
The all good clostridium sporogenes employment and suitability test (E & ST) bacterial strain of property, can be to avoid pathogenic risk.If there are clostridium sporogenes in drug, easily right
Patient causes infection etc., directly threatens the life of patient.The randomness and uncertainty that clostridium sporogenes is examined, are truly reflected inspection
Survey laboratory ability, this to improve Good Laboratory controlled level, ensure drug safety, or even break International trade practices,
Improving China's drug international competitiveness all has special important meaning.
Summary of the invention
Meet the quantitative life that employment and suitability test (E & ST) requires during drug microorganism detection the object of the present invention is to provide a kind of
Spore clostridium employment and suitability test (E & ST) bacterial strain；Meanwhile overcome the number of bacteria of work total in clostridium sporogenes employment and suitability test (E & ST) bacterial strain transport,
The problem of clump count can all change during storage and test etc., it is a further object to provide applicability examinations
Test the preparation method of bacterial strain, simple process, success rate height.
Present invention technical solution used for the above purpose is: clostridium sporogenes employment and suitability test (E & ST) bacterial strain, feature
Be: only comprising a kind of object bacteria: clostridium sporogenes, the aimed concn of object bacteria are 500~2000CFU/ branch, and every is equipped with 0.2g's
Freeze-dried powder.
Employment and suitability test (E & ST) bacterial strain in the sample category Medicine inspection is applied to applicability during drug microorganism detection
Test.
The employment and suitability test (E & ST) bacterial strain is using trehalose, gelatin and sterile water as matrix, and wherein trehalose volume fraction is
10%, gelatin volume fraction is 1%.
The preparation method of clostridium sporogenes employment and suitability test (E & ST) bacterial strain, it is characterized in that: including the preparation of sample addition bacterial strain, freeze-drying
Protectant preparation, sample freeze-drying, the uniformity of sample and five stability test, sample definite value steps, wherein sample is lyophilized
Process are as follows: bacterial strain recovery passage-increasing bacterium-bacterium solution packing-bacteria suspension preparation-freeze-drying and packaging-storage, detailed process are as follows:
(1) bacterial strain recovery passage
Reference culture is inoculated into clostridium enriched medium, so that it is recovered and is grown at 36 ± 1 DEG C, and to recovery
Bacterial strain is identified；
(2) increase bacterium
Object bacteria culture scrapes bacterium colony from inclined-plane, is added in the freeze drying protectant of 10mL and bacterium is made to the logarithmic growth end of term
Liquid；
(3) bacterium solution dispenses
The bacterium solution that a upper process obtains is mixed with freeze drying protectant, 100mL bacterium solution is prepared, this bacterium solution is placed in magnetic force and is stirred
Mix be stirred continuously on device it is lower be averagely dispensed into bacterium solution storage bottle, every bottle of 5mL, is placed in liquid nitrogen and saves backup by totally 20 bottles.
(4) bacteria suspension is prepared
According to target aimed concn 500~2000CFU/ branch of bacterium takes appropriate above-mentioned 5mL bacterium solution and 1500mL freeze drying protectant
Mixing, is placed on magnetic stirring apparatus to be stirred continuously down and is dispensed into sample bottle, 0.3mL/ bottles, in addition bottle stopper, but will there are gap,
Reality is not covered；
(5) it is lyophilized and packs
Sample bottle equipped with bacteria suspension is uncapped and is put into freeze dryer, 40~50h is freeze-dried, is tied when sample is freeze-dried
Shu Hou directly carries out jumping a queue in case, closing machine, and sample is vacuum state in sample bottle；
(6) it stores
It is kept in dark place under the conditions of sample is placed on -18 DEG C, random selected sample is issued to laboratory from whole samples
Or carry out uniformity and stability test.
The freeze drying protectant is the sterile water containing trehalose and skimmed milk power, and wherein volume fraction is respectively as follows: seaweed
Sugared 10%, gelatin 1%；The disposable bacterium solution for preparing same concentrations, the repeatable applicability for preparing multiple batches of same magnitude range
Test strain.
The uniformity of the sample and stability test are according to uniformity and stability test reference GB/T 15000.3-
2008 " rules and statistical method of standard sample work directive/guide (3) standard sample definite value " carry out.
Strain of the present invention is selected using clostridium sporogenes as object bacteria.
Clostridium sporogenes employment and suitability test (E & ST) bacterial strain of the invention has the advantage that characteristic: being a kind of with certain satisfaction detection
Method requires the employment and suitability test (E & ST) bacterial strain of levels, and sets about from sample stability, passes through the research of special process, stabilization
The research of property and uniformity, makes microorganism living, quantity not change, biochemical character does not morph, from meeting special fortune
Defeated condition forms a set of perfect employment and suitability test (E & ST) bacterial strain technology of preparing, is suitable for clostridium sporogenes employment and suitability test (E & ST) bacterial strain
Preparation.
Detailed description of the invention
Fig. 1 is process flow chart of the invention.
Specific embodiment
With reference to the accompanying drawing and specific embodiment present invention is further described in detail, but the invention is not limited to tools
Body embodiment.
Embodiment 1
Clostridium sporogenes employment and suitability test (E & ST) bacterial strain only includes a kind of object bacteria: clostridium sporogenes (Clostridium
Sporogenes), the aimed concn of object bacteria is 500~2000CFU/ branch, and every is equipped with the freeze-dried powder of 0.2g.The applicability
Test strain is using trehalose, gelatin and sterile water as matrix, and wherein trehalose volume fraction is 10%, gelatin volume fraction is
1%.
Employment and suitability test (E & ST) bacterial strain in the sample category Medicine inspection is applied to applicability during drug microorganism detection
Test.
Specific test process steps are as follows:
After sample is opened, 2mL sterile water is added immediately and carries out rehydration, the concussion that is vortexed mixes well, and 100 μ L is taken to be inoculated into
Fluid nutrient medium to be measured is applied on culture medium to be measured, 30~35 DEG C of 24~48h of culture.Whether good observe its growth,
Whether bacterium colony size, morphological feature are consistent.If well-grown, bacterium colony size, morphological feature are consistent, then illustrate tested culture medium and
Method is suitable for test product inspection, and the tested culture medium of on the contrary then explanation or method are not suitable for test sample inspection, should be according to method
Measure is further processed.
Embodiment 2
As shown in Figure 1, in a kind of embodiment 1 clostridium sporogenes employment and suitability test (E & ST) bacterial strain preparation method, including sample addition
The preparation of bacterial strain, the preparation of freeze drying protectant, sample freeze-drying, the uniformity of sample and stability test, sample definite value five steps
Suddenly, the specific steps are as follows:
1, the selection of sample addition bacterial strain
It only include a kind of object bacteria: clostridium sporogenes, the bacterial strain is purchased from specified mechanism, government, and demonstrate,proves with bacterial strain
Book ensure that the traceability of bacterial strain.
2, the preparation of freeze drying protectant
Sample is prepared as matrix (volume fraction: trehalose 10%, gelatin 1%) using trehalose, gelatin and sterile water and is lyophilized
Protective agent.
3, sample is lyophilized
(1) bacterial strain recovery passage
Reference culture is inoculated into clostridium enriched medium, so that it is recovered and is grown at 36 ± 1 DEG C, and to recovery
Bacterial strain is identified；
(2) increase bacterium
Object bacteria culture scrapes bacterium colony from inclined-plane, is added in the freeze drying protectant of 10mL and bacterium is made to the logarithmic growth end of term
Liquid；
(3) bacterium solution dispenses
The bacterium solution that a upper process obtains is mixed with freeze drying protectant, 50mL bacterium solution is prepared, this bacterium solution is placed in magnetic force and is stirred
Mix be stirred continuously on device it is lower be averagely dispensed into bacterium solution storage bottle, every bottle of 5mL, totally 10 bottles；
(4) bacteria suspension is prepared
According to target aimed concn 500~2000CFU/ branch of bacterium is prepared, and appropriate above-mentioned 5mL bacterium solution and 1500mL freeze-drying is taken to protect
Agent mixing is protected, is placed on magnetic stirring apparatus to be stirred continuously down and is dispensed into sample bottle (cillin bottle), 0.3mL/ bottles, in addition bottle stopper,
But reality is not covered there are gap；
(5) it is lyophilized and packs
Sample bottle equipped with bacteria suspension is uncapped and is put into freeze dryer, freeze drier parameter is by following setting:
Chilling rate (Cooling Rate) 0.5 DEG C/min
- 1 DEG C of 15min of early stage cold point (Incipient Freezing Point)
- 40 DEG C of cryogenic temperature (Cooling temperature)
- 29 DEG C of eutectic point (Melting Point Eutectic temperature)
20 DEG C of 120min of heating temperature (Heating temperature)
Start freeze drier, machine is directly entered the freezing dry process of sequencing, entire freeze-drying process 40h.
After sample freeze-drying, directly carry out jumping a queue in case；Closing machine.The not tight cillin bottle of plug is rejected,
Sample is vacuum state in cillin bottle；
(6) it stores
It is kept in dark place under the conditions of sample is placed on -18 DEG C, and the sample of preservation is suitably managed and detected, from complete
Random selected sample is issued to laboratory or carries out uniformity and stability test in portion's sample.
4, the uniformity and stability test of sample
Uniformity and stability inspection to object bacteria in sample are the main methods of verification sample preparation process validity.
Check sample homogeneity and stability according to uniformity and stability test according to GB/T 15000.3-2008 " standard sample work
Make the rule and statistical method of directive/guide (3) standard sample definite value " it carries out.Gained sample homogeneity and Detection of Stability method
And result is as follows:
12 samples are randomly selected respectively, using PetrifilmTMTotal plate count testing piece tests 2 × 12 in repeat condition
The clostridium sporogenes of part sample.Result data carries out statistical disposition with method of analysis of variance, and statistic procedure and result are as follows::
1 variance analysis formula of table
In upper table,
2 sample homogeneity testing result of table
3 sample homogeneity of table tests the results of analysis of variance
Conclusion: under 95% fiducial probability, compared with other factors are to the influence of test result, the inhomogeneities of sample is
It is acceptable.
Using two kinds of stability test: one is the stability test under storage temperature (4 DEG C), another kind is
Stability test at high temperature (traffic condition of analog sample), selects three temperature spots, respectively 20 DEG C, 36 DEG C and
42℃.Periodic detection sample tests 3 samples for different temperature points different holding time, by 2 × 3 parts of sample results
Mean value (after logarithmic transformed) carries out statistical disposition, F value < F with method of analysis of varianceCritical value, then illustrate the stabilization of employment and suitability test (E & ST) bacterial strain
Property meet the requirements, while determining and meeting longest holding time of employment and suitability test (E & ST) bacterial strain requirement under condition of different temperatures.Stablize
Property test result is shown in Table 4 and table 5.
4 sample short-term stability testing result of table
5 sample short-term stability of table tests the results of analysis of variance
5, sample definite value
Specific steps are as follows:
Using PetrifilmTMTotal plate count testing piece method, detects sample, according to GB/T 15000.3-2008
Corresponding principle carries out the meter of characteristic value in " rule and statistical method of standard sample work directive/guide (3) standard sample definite value "
Calculation and uncertain assessment.
Embodiment 3
Each step of the preparation method of clostridium sporogenes employment and suitability test (E & ST) bacterial strain described in the present embodiment is and in embodiment 2
Identical, different technical parameter are as follows: freeze-drying process 45h.
Embodiment 4
Each step of the preparation method of clostridium sporogenes employment and suitability test (E & ST) bacterial strain described in the present embodiment is and in embodiment 3
Identical, different technical parameter are as follows: freeze-drying process 50h.
Claims (6)
1. clostridium sporogenes employment and suitability test (E & ST) bacterial strain, it is characterized in that: only including a kind of object bacteria: clostridium sporogenes, the target of object bacteria
Concentration is 500~2000CFU/ branch, and every is equipped with the freeze-dried powder of 0.2g.
2. clostridium sporogenes employment and suitability test (E & ST) bacterial strain according to claim 1, it is characterized in that: in the sample category Medicine inspection
Employment and suitability test (E & ST) bacterial strain is applied to employment and suitability test (E & ST) during drug microorganism detection.
3. clostridium sporogenes employment and suitability test (E & ST) bacterial strain according to claim 1, it is characterized in that: the employment and suitability test (E & ST) bacterial strain with
Trehalose, gelatin and sterile water are matrix, and wherein trehalose volume fraction is 10%, gelatin volume fraction is 1%.
4. the preparation method of clostridium sporogenes employment and suitability test (E & ST) bacterial strain according to claim 1, it is characterized in that: adding including sample
Add the preparation of bacterial strain, the preparation of freeze drying protectant, sample freeze-drying, the uniformity of sample and stability test, sample definite value five
Step, wherein sample freeze-drying process are as follows: bacterial strain recovery passage-increasing bacterium-bacterium solution packing-bacteria suspension preparation-freeze-drying and packet
Dress-storage, detailed process are as follows:
(1) bacterial strain recovery passage
Reference culture is inoculated into clostridium enriched medium, so that it is recovered and is grown at 36 ± 1 DEG C, and to the bacterial strain of recovery
It is identified；
(2) increase bacterium
Object bacteria culture scrapes bacterium colony from inclined-plane, is added in the freeze drying protectant of 10mL and bacterium solution is made to the logarithmic growth end of term；
(3) bacterium solution dispenses
The bacterium solution that a upper process obtains is mixed with freeze drying protectant, 100mL bacterium solution is prepared, this bacterium solution is placed in magnetic stirring apparatus
On be stirred continuously it is lower be averagely dispensed into bacterium solution storage bottle, every bottle of 5mL, is placed in liquid nitrogen and saves backup by totally 20 bottles；
(4) bacteria suspension is prepared
According to target 500~2000 CFU/ branch of aimed concn of bacterium takes appropriate above-mentioned 5mL bacterium solution and 1500mL freeze drying protectant mixed
Close, be placed on magnetic stirring apparatus to be stirred continuously down and be dispensed into sample bottle, 0.3mL/ bottles, in addition bottle stopper, but will there are gaps, no
Cover reality；
(5) it is lyophilized and packs
Sample bottle equipped with bacteria suspension is uncapped and is put into freeze dryer, 40~50h is freeze-dried, when sample freeze-drying terminates
Afterwards, it directly carries out jumping a queue in case, closing machine, sample is vacuum state in sample bottle；
(6) it stores
Be kept in dark place under the conditions of sample is placed on -18 DEG C, from whole samples random selected sample be issued to laboratory or into
Row uniformity and stability test.
5. the preparation method of clostridium sporogenes employment and suitability test (E & ST) bacterial strain according to claim 4, it is characterized in that: the freeze-drying is protected
Shield agent is the sterile water containing trehalose and gelatin, and wherein volume fraction is respectively as follows: trehalose 10%, gelatin 1%；It is disposable to prepare
The bacterium solution of same concentrations, the repeatable employment and suitability test (E & ST) bacterial strain for preparing multiple batches of same magnitude range.
6. the preparation method of clostridium sporogenes employment and suitability test (E & ST) bacterial strain according to claim 4, it is characterized in that: the sample
Referring to GB/T 15000.3-2008, " standard sample work is led according to uniformity and stability test for uniformity and stability test
The then rule and statistical method of (3) standard sample definite value " it carries out.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110982763A (en) * | 2020-01-04 | 2020-04-10 | 广东环凯生物科技有限公司 | Stabilizer of clostridium perfringens and application thereof |
| CN110982762A (en) * | 2020-01-04 | 2020-04-10 | 广东环凯生物科技有限公司 | Clostridium sporogenes stabilizer and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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