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A method for quality control of bacterial vaccine strains

A technology for vaccines and strains, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as quality control of unseen bacterial vaccine strains

Active Publication Date: 2019-07-16
NAT INST FOR FOOD & DRUG CONTROL
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, with the development of biotechnology, a variety of molecular genetic analysis methods have come out one after another, such as pulse-field gel electrophoresis (PFGE) technology for the analysis of the whole genome of bacteria, but this method has not been used for bacterial vaccine strains. Therefore, in order to ensure the safety of the product from the source of the vaccine, it is urgent to establish a method for molecular genetic quality control of Leptospira vaccine strains to strengthen the quality control of vaccine strains

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  • A method for quality control of bacterial vaccine strains
  • A method for quality control of bacterial vaccine strains
  • A method for quality control of bacterial vaccine strains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 1

[0022] Example 1 General method

[0023] 1. Preparation of bacterial PFGE gel block

[0024] Take 10ml of Leptospira vaccine strain grown in rabbit serum phosphate medium at 28℃ for 7-10 days in a centrifuge tube, balance, centrifuge at 10000rpm 4℃ for 20min, discard the supernatant, and add 1ml buffer to the pellet (containing 1M Tris, 0.5M EDTA and 5M NaCl solution) were dissolved, transferred to a 1.5ml eppendorf tube, and centrifuged at 6000rpm for 5 minutes. Pour out the supernatant. According to the concentration of Leptospira, add 400μl buffer to dissolve the precipitate and mix well. Take 200μl of bacterial suspension and place it in a 1.5ml eppendorf tube, and incubate at 37°C for 5 minutes. Add the same amount of 1% gold medal agarose solution (purchased from Lonza, USA), mix well, add to the mold, and solidify at room temperature for about 15 minutes to obtain a bacterial PFGE gel block.

[0025] 2. Treatment of bacterial PFGE gel block

[0026] Take the bacterial PFGE g...

Embodiment 2

[0030] The following Leptospira vaccine strains (all from Wuhan Institute of Biological Products Co., Ltd.) were subjected to the PFGE analysis method as described in Example 1: Marker H9812; Jaundice haemorrhagic group (Lai strain); Autumn group (Lin 4 strains) ); Influenza typhoid fever group (Pro 6 strains); Marker H9812, obtained figure 1 The result shown.

[0031] This result indicates that the number, size and distribution of DNA fragments of different serogroups of Leptospira vaccine strains are different, showing different PFGE patterns.

Embodiment 3

[0033] Different generations of a Leptospira vaccine strain Influenza Typhoid Group (Pro 6 strains) (Leptospira Pro 6 strains before animal passage and the first generation of animal passages were from Wuhan Institute of Biological Products Co., Ltd.; then The Leptospira strains were passed on an in vitro artificial medium-rabbit serum phosphate medium every 7 days for a total of 20 generations.) Carry out the PFGE analysis method described in Example 1 to obtain figure 2 The result shown.

[0034] This result indicates that the influenza typhoid group (six strains) before the animal passage and 20 generations after the animal passage are completely consistent with the DNA bands of the bacterial species and have the same PFGE pattern. The culture characteristics and growth patterns of these different generation strains on rabbit serum phosphate medium are the same, and the serotype analysis results of the different generation strains are also consistent, which further supports t...

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Abstract

The invention relates to a leptospiral vaccine strain quality control method. The method comprises a, inoculating an animal with a leptospiral vaccine strain, collecting animal heart blood and culturing the leptospiras for leptospiral strain passage so that animal passage is realized, b, carrying out pulse-field gel electrophoresis (PFGF) analysis on different generations of the leptospiral vaccine strains before and after animal passage, and c, comparing the obtained DNA maps, wherein if the DNA maps are the same, different generations of the leptospiral vaccine strains before and after animal passage are stable. The method realizes analysis of the whole chromosome of the leptospiral strains and has the advantages of good repeatability, good distinguishing capability, stable result and clear result explanation.

Description

Technical field [0001] This application relates to the field of biological products, in particular to a method for quality control of bacterial vaccine strains. Background technique [0002] Strains for vaccine production refer to bacteria, viruses, spirochetes, rickettsia and other microorganisms isolated and screened from nature or clinically used for vaccine production and quality control. They are identified, classified and given fixed numbers. Pure culture of microorganisms. Strains are the material basis for vaccine research, production, and verification. Various bacterial vaccines, such as inactivated vaccines, live vaccines, or component-purified vaccines, are prepared through processes such as strain expansion. The quality of the production strain directly or indirectly affects the quality, safety and efficacy of the vaccine. Therefore, the strain must be strictly screened before it can be used as a vaccine production strain. At the same time, the production strain must...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
Inventor 王军志徐颖华张金龙辛晓芳
Owner NAT INST FOR FOOD & DRUG CONTROL
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