Clostridium difficile chromogenic medium and application thereof

A chromogenic medium, a technology for Clostridium difficile, applied in microorganism-based methods, microbial assay/inspection, microorganisms, etc., and can solve problems such as missed detection

Inactive Publication Date: 2016-08-10
ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The characteristics and identification of the colonies mainly depend on the visual observation and experience ju

Method used

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  • Clostridium difficile chromogenic medium and application thereof
  • Clostridium difficile chromogenic medium and application thereof
  • Clostridium difficile chromogenic medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1 standard bacterial strain test

[0019] 1. Components of chromogenic medium:

[0020]

[0021] Add 0.16 g of cycloserine, 0.07 g of cefoxitin sodium, the solvent is 1 L of deionized water, the amino acid is a mixture of proline and leucine at a mass ratio of 1:1, the pH value is 7.2, and it is used after pouring into a plate.

[0022] 2. Standard strains:

[0023] The standard strains of Clostridium difficile (ATCC700057, ATCC43255) purchased from the American Type Culture Collection (ATCC) were selected as positive controls, and Clostridium aerogenes ATCC13124, Proteus mirabilis ATCC12453, and Escherichia coli purchased from ATCC were used as positive controls. ATCC25922 was used as a negative control.

[0024] 3. Experimental operation

[0025] After recovery, the strains were inoculated into the chromogenic medium respectively, anaerobically cultured at 37±1°C for 18-24h, and the results were observed.

[0026] The results show that the standard s...

Embodiment 2

[0027] Embodiment 2 actual sample detection

[0028] 1. Components of chromogenic medium:

[0029]

[0030] Add 0.1 g of cycloserine, 0.15 g of cefoxitin sodium, the solvent is 1 L of deionized water, the amino acid is a mixture of proline and leucine at a mass ratio of 1:1, the pH value is 7.0, and it is used after pouring into a plate.

[0031] 2. Experimental operation

[0032] Take 200 mg of feces from patients with clinical diarrhea and mix them with 800 μl of absolute ethanol in a 1.5 ml centrifuge tube, cover the tube cap, and vortex to mix.

[0033] (2) The above samples were mixed and allowed to stand at room temperature for 1 hour.

[0034] (3) After standing still, put the centrifuge tube into a high-speed centrifuge and centrifuge at 800 rpm for 10 minutes. After centrifugation, the supernatant was aspirated and the pellet was retained.

[0035] (4) Add 800 μl of physiological saline to the precipitate and mix well.

[0036] (5) Pour all the mixed stool sam...

Embodiment 3

[0039] Embodiment 3 and the comparison of existing clostridium difficile selective plate

[0040] 1. Components of chromogenic medium:

[0041]

[0042]

[0043] Add 2.5 g of cycloserine, 0.01 g of cefoxitin sodium, the solvent is 1 L of deionized water, the amino acid is a mixture of proline and leucine at a mass ratio of 1:1, the pH value is 7.4, and it is used after pouring into a plate.

[0044] 2. Cycloserine-cefoxitin fructose agar (CCFA) medium was used as a control.

[0045] 3. Experimental operation

[0046] Take 200 mg of feces from patients with clinical diarrhea and 800 μl of absolute ethanol and mix them in 1.5 ml centrifuge tubes, cover the tube caps, and vortex to mix. Each sample was repeated once.

[0047] (2) The above samples were mixed and allowed to stand at room temperature for 1 hour.

[0048] (3) After standing still, put the centrifuge tube into a high-speed centrifuge and centrifuge at 800 rpm for 10 minutes. After centrifugation, the super...

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Abstract

The invention discloses a Clostridium difficile chromogenic medium and an application thereof. The chromogenic medium consists of 1-20g/L of tryptone, 1-20g/L of digested animal tissue, and 0.1-5.5g/L of glucose. L, yeast extract 0.2‑5.5g/L, sodium chloride 0.5‑15g/L, sodium sulfite 0.05‑1g/L, escin 0.1‑20g/L, iron 0.1‑5g/L, amino acid 0.2‑25g/L , agar powder 5-20g/L, cycloserine 0.1-2.5g/L and cefoxitin sodium 0.01-0.15g/L, the solvent is deionized water, pH value 7.0~7.6; the medium culture period of the present invention is 24 hours, Colonies are specifically black, and C. difficile isolates can be accurately obtained from clinical stool samples by color when using this medium.

Description

(1) Technical field [0001] The invention relates to a medium for cultivating Clostridium difficile, in particular to a chromogenic medium for cultivating Clostridium, which shortens the culture time and improves the detection rate and accuracy. (2) Background technology [0002] Clostridium difficile (Clostridium difficile) is a Gram-positive anaerobic bacillus, which is one of the normal flora in the human intestinal tract and is widely distributed in the natural environment and animal feces. It is reported that 25% to 30% of antibiotic-associated diarrhea is caused by Clostridium difficile, while pseudomembranous colitis is almost 100% caused by Clostridium difficile. C. difficile has seen a trend of increasing incidence, severity, and recurrence over the past two decades, all of which are associated with poor prognosis, and has become the number 1 "super bug" in Europe and the United States. Now the United States has Clostridium difficile infection and methicillin-resist...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12R1/145
CPCC12Q1/045
Inventor 罗芸
Owner ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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