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Culture medium for selectively separating and culturing group B streptococci and preparation method thereof

A technique for separating and culturing Streptococcus, applied in the field of pathogenic microbiology, can solve the problems of complex quorum sensing substances, difficult separation and purification, different effects, etc. Effect

Inactive Publication Date: 2017-02-22
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Quorum-sensing substances of Gram-positive bacteria that have been studied more at present are oligopeptides, which are specific to bacterial species and can promote bacterial metabolism. However, due to the similar molecular weight of such quorum-sensing substances, it is difficult to separate and purify, and the purification cost is high.
In addition, the quorum-sensing substances of bacteria are extremely complex. Artificially adding a single substance to the culture medium will have different effects under different conditions.

Method used

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  • Culture medium for selectively separating and culturing group B streptococci and preparation method thereof
  • Culture medium for selectively separating and culturing group B streptococci and preparation method thereof
  • Culture medium for selectively separating and culturing group B streptococci and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Comparative test of Columbia blood plate and group B streptococcus selective chromogenic basal medium

[0025] 1. Columbia blood plate was purchased from Antu Biotechnology Co., Ltd.

[0026] 2. Group B streptococcus selective chromogenic basal medium: glucose 2.5g, disodium diphosphate 1g, magnesium sulfate heptahydrate 0.41g, crystal violet 0.0002g, agar 3g, peptone 25g, soluble starch 20g, 4- Dissolve 11g of morpholine propanesulfonate, 8.5g of sodium dihydrogen phosphate, and 2g of yeast extract in 1L of distilled water, heat and mix, autoclave at 121°C for 25 minutes, cool to 60°C, and add through a bacterial filter (membrane filter) The sterilized antibiotics with a pore size of 0.22 μm: methotrexate 0.006g / L, colistin sulfate 0.005g / L, metronidazole 0.01g / L, poured onto a sterile plate, cooled at room temperature, and obtained group B chain Coccus selective chromogenic basal medium agar plate was set as the experimental group.

[0027] 3. Take the sp...

Embodiment 2

[0029] Embodiment 2: Comparative test of group B streptococcus selective chromogenic basal plate medium and group B streptococcus selective chromogenic basal plate medium added with culture supernatant

[0030] 1. Preparation of group B streptococcus selective chromogenic agar plates added with group B streptococcus culture supernatant

[0031] (1) Brain Heart Infusion Broth (BHI) liquid medium was used to inoculate and cultivate Group B Streptococcus (CM0109 strain) to the logarithmic growth phase, centrifuge at 10000g for 5 minutes, take the supernatant, and pass it through a bacterial filter (filter membrane pore size 0.22 μm) to obtain the culture supernatant.

[0032] (2) Group B Streptococcus selective chromogenic basal medium: glucose 2.5g, disodium diphosphate 1g, magnesium sulfate heptahydrate 0.41g, crystal violet 0.0002g, agar 3g, peptone 25g, soluble starch 20g, 4 - Morpholine propanesulfonate 11g, disodium hydrogen phosphate 8.5g, yeast extract 2g, dissolved in 1...

Embodiment 3

[0036] Embodiment 3: Common BHI medium and the BHI medium contrast test of adding culture supernatant

[0037] 1. Preparation of ordinary BHI medium: Weigh 38.5g of commercialized BHI dry powder (Qingdao Haibo Biotechnology Co., Ltd.), dissolve in 1L of distilled water, mix well, autoclave at 121°C for 25min, and dispense into L-shaped tubes Set as a control group.

[0038] 2. Preparation of BHI medium supplemented with culture supernatant

[0039] (1) Weigh 38.5 g of commercialized BHI dry powder (Qingdao Haibo Biotechnology Co., Ltd.), dissolve in 900 ml of distilled water, mix well, autoclave at 121 ° C for 25 min, add the culture supernatant obtained in Example 2 after cooling 100ml of solution was dispensed into L-shaped tubes as a control group.

[0040] 3. Inoculate 10ul of group B streptococci with McFarlane OD0.5, let it stand for 24 hours, and measure the OD every 4 hours. The experimental group enters the logarithmic growth phase faster than the control group, and...

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Abstract

The invention discloses a culture medium for selectively separating and culturing group B streptococci and a preparation method thereof. Culture supernatant fluid of group B streptococci at the logarithmic phase and an antibiotic combination are added to a group B streptococcus selective developing basic culture medium. The culture supernatant fluid contains quorum sensing substances capable of accelerating the growth of bacteria and facilitating the speed of color development reaction and color development intensity, thereby greatly shortening the detection time; and the antibiotic combination comprises amethopterin, colistin sulfate and metronidazole, which can inhibit the growth of mixed bacterium, except the group B streptococci, thereby achieving a purpose of selectively separating and culturing the group B streptococci. When the culture medium is used for detecting the group B streptococci, the characteristics of high specificity and sensitivity, convenience in observation and the like are provided, and the culture medium can be effectively applied to rapid detection to clinic samples.

Description

technical field [0001] The invention belongs to the field of pathogenic microbiology, and in particular relates to a culture medium for selectively isolating and cultivating group B streptococci and a preparation method thereof. Background technique [0002] Group B Streptococcus is a common opportunistic pathogen and a common pathogen that causes puerperal fever. These bacteria can cause puerperal infection no matter which way pregnant women give birth. Most of the puerperal infections caused by group B streptococci are single bacterial infections, with early onset of symptoms, high fever (>38.8°C), tachycardia, etc.; group B streptococci are also the main pathogens causing bacteremia and wound infection in pregnant women. Streptococcus pneumonia, meningitis, liver abscess and sepsis can also occur, and the mortality rate is relatively high. It has been reported that the mortality rate can reach 29% to 52%; Intrauterine infection can cause fetal death, but the most comm...

Claims

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Application Information

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IPC IPC(8): C12Q1/14C12Q1/04C12R1/46
CPCC12Q1/14C12Q1/045
Inventor 玄英花邓超
Owner JIANGNAN UNIV
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