Cronobacter broth culture medium and detection method

A technology of Cronobacter, broth culture medium, applied in biochemical equipment and methods, microbial determination/inspection, fluorescence/phosphorescence, etc., can solve problems such as time-consuming, shorten time, improve specificity and accuracy Sexual, low-cost effects

Inactive Publication Date: 2011-12-21
CHINESE ACAD OF INSPECTION & QUARANTINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] As can be seen, existing assays and assay media require separate enrichment and testing steps and are therefore time consuming

Method used

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  • Cronobacter broth culture medium and detection method
  • Cronobacter broth culture medium and detection method
  • Cronobacter broth culture medium and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Selective evaluation of the Cronobacter bouillon medium of the present invention

[0036] 1. Medium Preparation

[0037] Weigh each component according to the formula in Table 1, and prepare three kinds of culture medium, namely medium 1, medium 2 and medium 3, adjust the pH of medium 1 and medium 2 to 7.4±0.1; adjust the culture medium The pH of base 3 was pH 6.8±0.2, sterilized by autoclaving at 121°C for 15 minutes, cooled to room temperature, and dispensed into test tubes, each with 10 mL for use.

[0038] Prepare TSB broth medium (product of American BD Company, product number: 211825) according to the manufacturer's instructions, and set aside.

[0039] Table 1: Media Components

[0040]

[0041] ** 10 mg of vancomycin was dissolved in 10 mL of distilled water, sterilized by filtration, and added to the autoclaved medium.

[0042] 2. Inoculation

[0043] Inoculate the reference strain of Cronobacter and related reference strains in TSB broth med...

Embodiment 2

[0050] Embodiment 2: the specificity test of the Cronobacter bouillon medium of the present invention

[0051] In order to further verify the specificity and selectivity of the Cronobacter bouillon medium of the present invention, reference strains of all 6 species of Cronobacter and 5 isolated strains were selected. The most closely related E. aerogenes, E. cloacae and other strains that are more common but microbiologically distant (see Table 2) were tested below.

[0052] 1. Medium Preparation

[0053] According to the formula of medium 2 in Table 1, each component was weighed to prepare the medium, and after sterilization, it was cooled to 50° C., and divided into test tubes, each of which was 10 mL for use.

[0054] 2. Inoculation

[0055] With Example 1, the representative strains in Table 2 were selected and inoculated in TSB broth medium (product of BD Company in the United States, article number: 211825), cultivated at 36° C. for 18-24 h, and a circle was streaked a...

Embodiment 3

[0061] Example 3: The optimum culture temperature of the culture medium of the present invention

[0062] 1. the preparation of Cronobacter bouillon medium of the present invention

[0063] Prepare according to the formulations of media 3 and 4 in Table 3. The specific preparation method is as described in Example 1.

[0064] Table 3: Medium components for optimum culture temperature testing

[0065] Element

source of material

Medium 3

Medium 4

Mixed peptone (g)

Beijing Aoboxing

20

×

Gelatin peptone (g)

Beijing Aoboxing

×

10

Sodium chloride (g)

Sigma

34

×

Lactose (g)

Sigma

5

×

Glucose (g)

Sigma

×

5

α-MUG (g)

Sigma (Cat. No.: M9766, CAS: 17833-43-1)

0.08

0.09

Sodium Lauryl Sulfate (g)

Sigma

0.1

×

Bile salt (g)

Sigma

×

20

Bright green (mg)

Sigma

×

10mg

...

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Abstract

The invention relates to a detection medium and a detection method for Cronobacter in food. The detection method of the present invention utilizes the activity of alpha-glucosidase produced by Cronobacter to detect Cronobacter in liquid culture medium, and can identify Cronobacter while carrying out selective enrichment. The Cronobacter broth medium of the present invention may comprise 18-20 parts by weight of mixed peptone; 3-5 parts by weight of lactose; 2-3 parts by weight of K2HPO4; 2-3 parts by weight of KH2PO4; 20-34 parts by weight of sodium chloride 0.08-0.15 parts by weight of sodium lauryl sulfate; 0.009-0.011 parts by weight of vancomycin; 0.07-0.10 parts by weight of 4-methylumbelliferone-α-D-glucopyranoside; pH6.8 ± 0.2, the best Culture temperature, 36°C. The invention also discloses a reagent kit for the above detection method. The invention has the advantages of shortening the detection time and avoiding subsequent tedious identification operation steps, thereby improving the detection efficiency of Cronobacter.

Description

technical field [0001] The invention relates to a Cronobacter broth broth culture medium and a detection method, belonging to the field of food sanitation microbial safety inspection and monitoring. Background technique [0002] Cronobacter spp., formerly known as Enterobacter sakazakii, widely exists in nature and can cause neonatal meningitis, sepsis, necrotizing colitis, and infection in infants and young children through formula milk powder and other foods The fatality rate is as high as 20% to 50%, which has attracted the attention of the World Health Organization, governments of various countries and experts in the food industry. This type of bacteria was defined and classified as a species of Enterobacter in 1980, namely Enterobacter sakazakii, and was reclassified as a new genus in 2008, namely Cronobacter, which included 5 new species, 1 A Cronobacter genotype (genomospeciese) and 3 new subspecies (Zhao Guiming et al., Taxonomic changes and biological characteristi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/10G01N21/64
Inventor 赵贵明高旗利陈颖
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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