Enterobacter sakazaii colour development culture medium, detection kit and detection method

A technology of Enterobacter sakazakii and chromogenic medium, which is applied in the field of detection methods and compositions used, can solve the problems of high detection cost, high technical requirements, complicated equipment operation, etc. Sensitive effect

Inactive Publication Date: 2009-05-06
GUANGDONG HUANKAI MICROBIAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these technologies also have certain defects, such as high technical requirements, complicated equipment operation, professional technicians are required to operate, and expensive testing costs, etc.

Method used

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  • Enterobacter sakazaii colour development culture medium, detection kit and detection method
  • Enterobacter sakazaii colour development culture medium, detection kit and detection method

Examples

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Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Verification of the specificity of Enterobacter sakazakii chromogenic medium according to the present invention

[0021] 1. Preparation of Enterobacter sakazakii chromogenic medium: Weigh 39.62g of Enterobacter sakazakii chromogenic medium (containing 11g of peptone, 5g of beef extract powder, 5g of sodium chloride, 1.5g of bile salt, 15g of agar, chromogenic medium Substrate X-a-glucopyranoside (Apoloscientific, BIMB1190) 0.1g, Na 2 CO 3 0.02g, 1.0g ferric ammonium citrate, 1.0g sodium thiosulfate), add 1000mL distilled water or deionized water, stir, heat and boil until completely dissolved, autoclave at 121℃ for 15min, cool to 40~50℃, pour plate ,spare;

[0022] 2. Inoculation: Enterobacter sakazakii, Proteus vulgaris, P. mirabilis, Pseudomonas aeruginosa, Escherichia coli, Salmonella typhimurium , Serralia marcescens, Streptococcus faecalis, and Staphylococcus aureus were resuscitated on nutrient agar for 24 hours, respectively streaked and inoculated...

Embodiment 2

[0024] Example 2: Verification of sensitivity and specificity of Enterobacter sakazakii chromogenic medium according to the present invention

[0025] 1. Preparation of Enterobacter sakazakii chromogenic medium: Weigh 39.62g of Enterobacter sakazakii chromogenic medium (containing 11g of peptone, 5g of beef extract powder, 5g of sodium chloride, 1.5g of bile salt, 15g of agar, chromogenic medium Substrate X-a-glucopyranoside (Apoloscientific, BIMB1190) 0.1g, Na 2 CO 3 0.02g, 1.0g ferric ammonium citrate, 1.0g sodium thiosulfate), add 1000mL distilled water or deionized water, stir, heat and boil until completely dissolved, autoclave at 121℃ for 15min, cool to 40~50℃, pour plate ,spare;

[0026] 2. Inoculation: Enterobacter sakazakii ATCC51329, HK7402, Proteus vulgaris CMCC (B) 49027, Proteus mirabilis CMCC (B) 49005, Pseudomonas aeruginosa ATCC9027, ATCC27853, Escherichia coli ATCC25922, Salmonella typhimurium CMCC (B) 50115, Serratia marcescens HK7121, Enterococcus faecal...

Embodiment 3

[0038] Example 3: Detection of Enterobacter sakazakii in artificially contaminated samples

[0039] 1. Preparation of chromogenic plate

[0040] Weigh 39.62g of chromogenic medium dry powder (containing peptone 11g, beef extract powder 5g, sodium chloride 5g, bile salt 1.5g, agar 15g, chromogenic substrate X-a-glucopyranoside (Apoloscientific, BIMB 1190) 0.1g, Na 2 CO 3 0.02g, 1.0g ferric ammonium citrate, 1.0g sodium thiosulfate), add 1000mL distilled water or deionized water, stir, heat and boil until completely dissolved, autoclave at 121℃ for 15min, cool to 40~50℃, pour plate ,spare;

[0041] 2. Pre-enrichment bacteria

[0042] Weigh 8 parts of each of the 4 milk powder samples, each 100g, and add 4 samples to the triangular flask containing 900mL BP enrichment solution by aseptic operation, 3 of which are added with three concentrations of Enterobacter sakazakii; the other 4 The samples were added to Erlenmeyer flasks containing 900mL of sterile water, three of which...

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Abstract

The invention provides a fluorogenic culture medium of Enterobacter sakazakii, a test kit and a test method thereof, and relates to a method for testing microbe and a composition used by the method. The fluorogenic culture medium comprises the compositions as follows: 8 to 20 grams of peptone, 4 to 7 grams of beef extract powder, 4 to 7 grams of sodium chloride, 1.0 to 2.0 grams of cholate, 12 to 20 grams of agar, 0.05 to 0.5 grams of X-a-glucopyranoside, 0.01 to 0.04 grams of Na2CO3, 0.3 to 2.0 grams of ferric ammonium citrate, and 0.3 to 2.0 grams of sodium thiosulfate. The test kit consists of the fluorogenic culture medium of Enterobacter sakazakii, an enriched liquid A ,namely buffer peptone water culture medium, and an enriched liquid B, namely modified lauryl sulfate pancreas peptone broth. The test kit is configured simply, and the test method has high sensitivity in detection, so the method is suitable for treating large flux samples.

Description

【Technical field】 [0001] The invention relates to a method for detecting microorganisms and a used composition. 【Background technique】 [0002] Food microbiological safety is the top priority in food safety, and Enterobactersakazakii is a newly discovered pathogen in dairy products that has attracted widespread attention. In 1961, two British scientists reported two cases of meningitis caused by Enterobacter sakazakii for the first time, and then successively reported neonatal Enterobacter sakazakii infection in the United States, Greece, the Netherlands, Iceland, Canada, Belgium and other countries. It can be seen that sporadic and outbreak cases of infants, premature infants, meningitis, sepsis and necrotizing colitis caused by Enterobacter sakazakii have appeared one after another around the world. Fatal cases can be as high as 40% to 50%. A number of research reports have shown that infant formula milk powder is the main infection channel for infants, premature infants...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04G01N21/78
CPCY02A50/30
Inventor 卢勉飞蔡芷荷吴清平
Owner GUANGDONG HUANKAI MICROBIAL SCI & TECH
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