Listeria enriched culture medium and preparation method

A Listeria and culture medium technology, applied in the field of microbiology, can solve the problems of the existence of non-pathogenic Listeria and other environmental bacteria, hidden dangers to human health, cumbersome and time-consuming problems, and achieve shortened culture time, improved sensitivity, and damage The effect of diminishing

Active Publication Date: 2017-09-08
ICDC CHINA CDC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the EN ISO isolation method can isolate and culture lower amounts of pathogenic Listeria in the sample, non-pathogenic Listeria and other environmental bacteria can also be present in the sample
Since non-pathogenic Listeria and other environmental bacteria have better adaptability to the enrichment medium, in the process of enrichment culture of the enrichment medium, the non-pathogenic Listeria and other environmental bacteria are too fast Growth can mask the presence of pathogenic Listeria, resulting in false negative test results and serious human health concerns
In addition, the current method of isolating and cultivating Listeria requires a two-step enrichment process, which is very tedious and time-consuming.

Method used

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  • Listeria enriched culture medium and preparation method
  • Listeria enriched culture medium and preparation method
  • Listeria enriched culture medium and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Comparison of utilization of biochemical substrates by pathogenic Listeria and harmless Listeria

[0030] 1. Strains

[0031] Listeria monocytogenes reference strain (ATCC-BAA-678, EGD-e), Listeria harmless reference strain (ATCC-BAA-680, LIN), Listeria eziri reference strain (ATCC-BAA-679, PAM).

[0032] 2. Reagents

[0033] Reagents required for BIOLOG experiments, including magnesium chloride, calcium chloride, arginine, glutamic acid, L-cystine, 5'-uridine sodium, yeast extract, Tween-80, double distilled water, BIOLOG color development Dye Dye mix-F.

[0034] 3. Culture medium preparation

[0035] Magnesium chloride, calcium chloride, arginine, glutamic acid, L-cystine, sodium 5'-uridine, yeast extract, Tween-80, and double distilled water were prepared as additives according to the operating procedures provided by BIOLOG. Then the additives were mixed with the culture solution IF-0aGN / GP provided by BIOLOG company and the chromogenic dye Dye mix-F ...

Embodiment 2

[0044] Embodiment 2. Design and preparation of basic enrichment medium

[0045] According to the experimental results of Example 1, β-D-allose was used as the carbohydrate of the medium, and on the basis of adding other nutrients and chemical components, the following basal medium formula was designed and prepared:

[0046]

[0047]

[0048] Dissolve the above components in 1000ml distilled water, adjust the pH value to 7.2, mix well, sterilize at 121°C for 15min, cool and store at 4°C for later use.

Embodiment 3

[0049] Example 3. The growth rate comparison of bacteria in the basic enrichment medium

[0050] 1. Strains

[0051] Listeria monocytogenes reference strain (ATCC-BAA-678, EGD-e), Listeria harmless reference strain (ATCC-BAA-680, LIN), Listeria eziri reference strain (ATCC-BAA-679, PAM).

[0052] 2. Reagents

[0053] With embodiment 2.

[0054] 3. Culture medium preparation

[0055] Prepared according to Example 2.

[0056] 4. Bacterial Culture

[0057] Listeria monocytogenes reference strain (ATCC-BAA-678, EGD-e), Listeria harmless reference strain (ATCC-BAA-680, LIN), Listeria eziri reference strain (ATCC-BAA-679, PAM) was cultured in liquid brain-heart medium to OD=0.6, and then diluted to a specific concentration with phosphate buffered saline (PBS).

[0058] 5. Growth Rate Determination

[0059] Inoculate the same amount of Listeria monocytogenes, Listeria eziri, and Listeria innocua into three OD tubes containing 5ml of allose-based enrichment medium, and take ou...

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Abstract

The invention discloses a culture medium used for performing selective separation on pathogenic listeria. The culture medium comprises base components including tryptone, peptone, beef extract powder, sodium chloride and disodium hydrogen phosphate, the culture medium also comprises beta-D-allose; more preferably, the culture medium also comprises acid trypaflavine, nalidixic acid and lithium chloride. Compared with a traditional enriched culture medium, the culture medium provided by the invention is increased in sensitivity by 16 percent when separating the pathogenic listeria, and a culture step is reduced, and 48 hours of culture time is shortened.

Description

technical field [0001] The invention discloses a selective bacteria-enrichment medium, which belongs to the field of microbiology. Background technique [0002] Listeria monocytogenes are facultative intracellular gram-positive bacteria. At present, there are 17 species of Listeria genus, among which the pathogenic Listeria include Listeria monocytogenes and Listeria iridii, which can cause listeriosis in humans and animals, and are important food-borne human and animal diseases. comorbid pathogens. The clinical manifestations are meningitis in humans and animals, sepsis, abortion in pregnant women, febrile gastroenteritis, etc., and the clinical mortality rate is relatively high. The bacterium widely exists in soil, meat products and aquatic products, etc., and can survive in various conditions such as high salt, low temperature, and dryness, and is mainly spread through contaminated food. Since the 1980s, Listeria monocytogenes has repeatedly broken out in European and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12R1/01
CPCC12Q1/045
Inventor 叶长芸刘东鑫王艳
Owner ICDC CHINA CDC
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