Color developing culture medium, detecting kit and detecting method for vibrio parahaemolyticus

A hemolytic Vibrio and chromogenic culture medium technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection, can solve the problem of inability to distinguish live bacteria from dead bacteria and difficult to meet detection needs , Expensive testing costs and other issues, to achieve the effects of saving testing costs and time, short cycle time, and simple configuration

Inactive Publication Date: 2007-06-06
广东实验中学 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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.
The PCR method needs to enrich bacteria first, and cannot distinguish live bacteria from dead bacteria, which is prone to false positives; immunological methods also need to prepare high-purity antibodies, so these technologies are difficult to meet the needs of enterprises and government regulators for the detection of a large number of food samples
[0005] At present, French Chemagar company has developed a chromogenic medium for Vibrio parahaemolyticus, but the price is expensive, and the supporting products and reagents need to be imported, and the detection cost is very high. Therefore, some basic-level detection institutions in China seldom use it.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Verification of the specificity of Vibrio parahaemolyticus chromogenic medium of the present invention

[0022] 1. Preparation of Vibrio parahaemolyticus chromogenic medium: Take 15g of peptone, 3g of yeast powder, 10g of NaCl, 20g of sucrose, 0.11g of monosaccharide, 10g of sodium citrate, 10g of sodium thiosulfate, 8g of bile salt, mix Chromogenic substrate Y-glucoside (US sigma company) 0.35g, agar 12g, NaCO3 1g, water 1000mL, mix and heat until the agar is completely dissolved, boil for 1-2min, adjust the pH to 8.6±0.2, wait to cool to 40-50°C , pour into a flat plate and set aside.

[0023] 2. Inoculation: Vibrio parahaemolyticus (Vibrio parahaemolyticus), Vibrio cholerae (Vibriocholera), Vibrio alginolyticus (Vibrioalginolyticus), Vibrio vulnificus (Vibrio vulnificus), Vibrio mimicus (Vibrio mimicus) in 3.5% NaCl nutrition Agar, Eschetichiacoli (Eschetichiacoli), Salmonella (Salmonella) on nutrient agar, Listeria monocytogenes (Listeriamonocytogenes...

Embodiment 2

[0025] Embodiment 2: Verification of sensitivity and specificity of Vibrio parahaemolyticus chromogenic medium according to the present invention

[0026] 1. Preparation of Vibrio parahaemolyticus chromogenic medium: Take 15g of peptone, 3g of yeast powder, 10g of NaCl, 20g of sucrose, 0.11g of monosaccharide, 10g of sodium citrate, 10g of sodium thiosulfate, 8g of bile salt, mix Chromogenic substrate Y-glucoside (US sigma company) 0.35g, agar 12g, NaCO3 1g, water 1000mL, mix and heat until the agar is completely dissolved, boil for 1-2min, adjust the pH to 8.6±0.2, wait to cool to 40-50°C , pour into a flat plate and set aside.

[0027] 2. Vaccination:

[0028] After resuscitating Vibrio parahaemolyticus in 3.5% NaCl nutrient agar for 24 hours, pick 1 ring from the inoculation loop, add it to 10mL 0.85% normal saline to make stock solution, and then carry out 10-fold gradient dilution, take 10 -4 -10 -6 Concentration of 1mL bacterial solution, respectively, coated the abov...

Embodiment 3

[0034] Embodiment 3: the comparison of vibrio parahaemolyticus detection method of the present invention and national standard detection method (abbreviation national standard method)

[0035]Collect a large amount of marine products, freshwater fish etc. from market, adopt national standard method (GB / T4789.7-2003, food sanitation microbiological inspection Vibrio parahaemolyticus inspection) and detection method of the present invention to Vibrio parahaemolyticus respectively The detection was carried out, and the positive samples were verified with the French Mérieux Bacteria Identification System-API reagent strip, and the sensitivity of the two detection methods was compared.

[0036] (1) National standard method: sample—sodium chloride polymyxin B broth (or sodium chloride crystal violet) enriched for 24 hours—streaked inoculation on TCBS plate—blue-green colony—biochemical identification.

[0037] (2) Detection method of the present invention: sample-contain 3.5%NaCl TS...

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Abstract

The present invention relates to color developing culture medium, detecting kit and detection method for Vibrio parahaemolyticus, and belongs to the field of food microbe safety monitoring technology. The detecting kit consists of color developing culture medium comprising bacterial specific enzyme in 0.05-0.95 g and mixed color developing substrate Y-glucoside, enrichment liquid A of pancreatic casitone liquid culture medium and enrichment liquid B comprising sodium chloride, polymyxin and broth. The detection method includes the enrichment culture of the sample in enrichment liquid A and enrichment liquid B, the culture of the two parts of enrichment cultured liquid in the color developing culture medium, and judging whether to exist Vibrio parahaemolyticus according to whether to have smooth raised purple colony of 2-3 mm diameter. The present invention has low cost, high detection sensitivity, short detection period and other advantages.

Description

【Technical field】 [0001] The present invention relates to a microbial chromogenic culture medium, a detection kit and a detection method, in particular to a chromogenic culture medium, a detection kit and a detection method of Vibrio parahaemolyticus, belonging to the field of food microbial safety monitoring. 【Background technique】 [0002] "Food is the most important thing for the people", food is the material basis for human survival, and food safety is a major issue related to human health and social development. In recent years, vicious food safety incidents at home and abroad have occurred continuously, especially foodborne diseases caused by Salmonella, enterohemorrhagic Escherichia coli, Listeria monocytogenes, and Vibrio parahaemolyticus. According to statistics, about 1 / 3 of people in developed countries suffer from food-borne diseases every year, and there are about 76 million cases of food-borne diseases in the United States every year. Food-borne diseases have b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12R1/63
Inventor 曾婧张淑红张菊梅傅岚
Owner 广东实验中学
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