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Bacillus subtilis applicability test strain and preparation method thereof

A Bacillus subtilis, applicability technology, applied in the field of Bacillus subtilis applicability test strains and its preparation, can solve the problems of threatening the life of patients and infection of patients

Pending Publication Date: 2019-06-11
浙江宝录检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If there is Bacillus subtilis in the medicine, it is very easy to cause infection to the patient, directly threatening the life of the patient

Method used

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  • Bacillus subtilis applicability test strain and preparation method thereof
  • Bacillus subtilis applicability test strain and preparation method thereof
  • Bacillus subtilis applicability test strain and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The Bacillus subtilis applicability test strain contains only one target bacteria: Bacillus subtilis, and the target concentration of the target bacteria is 500-2000 CFU / piece, and each piece contains 0.2g of freeze-dried powder. The applicability test strain is based on trehalose, skimmed milk powder and sterile water, wherein the volume fraction of trehalose is 10% and the volume fraction of gelatin is 1%.

[0030] The sample belongs to the strain used for the applicability test in drug inspection, and is applied to the applicability test in the process of drug microorganism inspection. The specific test process steps are as follows:

[0031] After the sample is opened, immediately add 2 mL of sterile water for rehydration, vortex and shake to mix well, take 100 μL to inoculate the liquid medium to be tested or spread it on the medium to be tested, and incubate at 30-35°C for no more than 3 days. sky. Observe whether the growth is good, and whether the colony size a...

Embodiment 2

[0033] like figure 1 As shown, a preparation method of the Bacillus subtilis applicability test strain in Example 1, including the preparation of the sample addition strain, the preparation of the freeze-drying protective agent, the freeze-drying of the sample, the uniformity and stability test of the sample, the sample determination Five steps, the specific steps are as follows:

[0034] 1. Selection of sample addition strains

[0035] It contains only one target bacteria: Bacillus subtilis, all of which are purchased from government-designated institutions and are accompanied by a strain certificate to ensure the traceability of the strains.

[0036] 2. Preparation of lyophilized protective agent

[0037] The samples were prepared with trehalose, gelatin and sterile water as the matrix (volume fraction: trehalose 10%, gelatin 1%) to prepare a lyoprotectant.

[0038] 3. Sample freeze-drying

[0039] (1) Strain recovery and passage

[0040] Inoculate the standard strains ...

Embodiment 3

[0080] The steps of the preparation method of the Bacillus subtilis applicability test strain described in this example are the same as those in Example 2, and the different technical parameters are: the freeze-drying process is 45h.

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Abstract

The invention belongs to the field of quality control in drug microbiological detection, and particularly relates to a bacillus subtilis applicability test strain and a preparation method thereof. Thebacillus subtilis applicability test strain only contains a certain amount of bacillus subtilis. The uniformity and the stability meet the requirements of the strain for the applicability test, the bacterial count of the strain is within the standard requirement range of the applicability test, the requirements of the applicability test, the growth promotion test and the like in a drug microorganism detection process are met to the maximum extent, and the sample preparation method is simple in process and high in success rate.

Description

technical field [0001] The invention belongs to the quality control field of microbiological detection, and particularly relates to a Bacillus subtilis applicability test strain and a preparation method thereof. Background technique [0002] Due to the particularity of drugs, the field of drug microbiology testing is very special, and it is required that the test materials such as culture medium must be checked for suitability before each test, and there are requirements for the concentration range of the test strains. Most of the existing pure strains have not been quantified and can only be used for qualitative use, and there are only a few quantitative strains. For this type of quantitative strain, uniformity and stability are particularly important. The existing quantitative strains have problems such as poor stability and high requirements for transportation conditions. [0003] Bacillus subtilis belongs to the genus Bacillus and is a Gram-positive aerobic bacterium th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/125
Inventor 杨洁王倩赵红阳谢艳王珏
Owner 浙江宝录检测技术有限公司
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