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A culture medium for enriching ureaplasma, a protective agent for freeze-drying and preserving ureaplasma, a preparation method thereof, and a method for freeze-drying and preserving ureaplasma

A technology of ureaplasma and culture medium, applied in the field of microorganisms, can solve problems such as poor preservation effect and difficulty in enriching ureaplasma, and achieve the effects of reducing the difference between batches, avoiding frequent preservation, and expanding the number of preservations

Active Publication Date: 2021-09-24
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of this, the object of the present invention is to provide a culture medium for enriching Ureaplasma and a protective agent for freeze-drying and preserving Ureaplasma for the problems of difficulty in enriching Ureaplasma and poor preservation effect in the prior art. And freeze-drying method for preserving ureaplasma

Method used

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  • A culture medium for enriching ureaplasma, a protective agent for freeze-drying and preserving ureaplasma, a preparation method thereof, and a method for freeze-drying and preserving ureaplasma
  • A culture medium for enriching ureaplasma, a protective agent for freeze-drying and preserving ureaplasma, a preparation method thereof, and a method for freeze-drying and preserving ureaplasma
  • A culture medium for enriching ureaplasma, a protective agent for freeze-drying and preserving ureaplasma, a preparation method thereof, and a method for freeze-drying and preserving ureaplasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Comparing the enrichment effect of different culture media for Ureaplasma

[0024] Taking Ureaplasma urealyticum ATCC33175 as an example, the enrichment effect of several different media for Ureaplasma urealyticum was compared. The culture medium involved in the present invention is marked as A, and is respectively marked as A1, A2, A3, and A4 according to the different amounts of each component in the formula. The culture medium involved in CLSI M43-A (Page 24) in the United States is marked as B, the culture medium involved in "Practical Clinical Microbiology Test and Atlas" (Page 120) is marked as C, and the "Microbial Culture Medium Quality Control and Diagram" (Page 236 ) The culture medium involved is marked as D.

[0025] Wherein, the formulation of medium A is shown in Table 1.

[0026] Formulation and preparation method of medium A of table 1

[0027]

[0028]

[0029] Note: The above formula is the formula dosage for a total of 1L medium. ...

Embodiment 2

[0039] Example 2. Comparing the enrichment effect of different methods of adding protective agent on ureaplasma

[0040] Taking Ureaplasma urealyticum ATCC33175 as an example, the effects of different addition methods of protective agent on the concentration of Ureaplasma urealyticum after freeze-drying were compared.

[0041] Prepare medium A1 according to the formula and method of Example 1.

[0042] Preparation of protective agent: anhydrous trehalose 100g L -1 , Tryptone 50g·L -1 , Polyvinylpyrrolidone K30 20g·L -1 , sterilized at 121°C for 15 minutes, part of which was placed in a sterile environment for use, and part of which was divided into 1 mL / bottle under aseptic conditions and then freeze-dried for use.

[0043] Inoculate 50 μL of Ureaplasma urealyticum ATCC33175 strain into about 100 mL of the above-mentioned medium A, culture at 37°C for 14 hours, take 100 μl in a sterile environment every 30 minutes, put it in a glass bottle, and measure it with a micro-acidi...

Embodiment 3

[0049] Embodiment three, the preservation effect comparison of different ureaplasma

[0050] 4 strains of different Ureaplasma were preserved by the preservation method described in the present invention. The 4 strains of Ureaplasma were U. urealyticum ATCC33175, ATCC27618, U. minutica ATCC27813, ATCC700970. The 4 strains of Ureaplasma were preserved in three batches.

[0051] Prepare medium A1 according to the formula and method of Example 1.

[0052] Preparation of protective agent: anhydrous trehalose 100g.L -1 , Tryptone 50g.L -1 , Polyvinylpyrrolidone K30 20g.L -1 , sterilized at 121°C for 15 minutes, divided into 1mL / bottles under sterile conditions, and freeze-dried for later use.

[0053] Inoculate 50μL-100μL of 4 strains of Ureaplasma strains into about 100mL of the above-mentioned medium A, culture them at 37°C for 14 hours, take 100μl in a sterile environment every 30min, put them in a glass bottle, and use a micro-acidity meter Measure its pH value until its p...

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Abstract

The application belongs to the field of microorganisms, and discloses a culture medium for enriching ureaplasma, a protective agent for freeze-drying and preserving the ureaplasma, a preparation method thereof, and a method for freeze-drying and preserving the ureaplasma. The medium for enriching ureaplasma in the present invention has stronger buffer capacity and is more conducive to the enrichment of ureaplasma. The protective agent of the present invention is relatively stable, which reduces the risk of contamination of the ureaplasma during storage, and can expand the number of storage and reduce labor costs. The protective agent of the present invention is applied to the preservation of ureaplasma, and the stability of ureaplasma is greatly improved. The method for freeze-drying and preserving Ureaplasma in the present invention can monitor the growth state of Ureaplasma by measuring the pH of the bacterial suspension, which is more intuitive and quantifiable, and is conducive to maintaining stability between different batches. Further use the pre-lyophilized protective agent to freeze-dry and store Ureaplasma. The two-time freeze-drying storage method will not dilute the Ureaplasma suspension, which can enrich it to the greatest extent and effectively improve the freeze-drying process. after concentration.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a method for freeze-drying and preserving ureaplasma, in particular to a culture medium for enriching ureaplasma, a protective agent for freeze-drying and preserving ureaplasma, a preparation method thereof and a method for freeze-drying and preserving ureaplasma Primitive method. Background technique [0002] Ureaplasma is an independent new genus separated from Mycoplasma, including six species, among which Ureaplasma urealyticum (M.urealyticum) is pathogenic. Ureaplasma urealyticum, also known as Ureaplasma urealyticum, is one of the six genus Ureaplasma, which is closely related to human genitourinary tract infections and is one of the common parasites in the human genitourinary tract. environment can cause disease. Ureaplasma urealyticum can use its own urease to decompose urea to provide energy for metabolism. The colonization of Ureaplasma urealyticum in the hu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/04C12R1/01
CPCC12N1/04C12N1/20
Inventor 吴旭晓王则宇杨洋刘亚锋付光宇杨增利
Owner AUTOBIO DIAGNOSTICS CO LTD
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