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Quick detecting coliform group and medium for culturing coliforms and preparation method

A technology of coliform bacteria and Escherichia coli, which is applied in the field of health and epidemic prevention detection and environmental water quality monitoring, can solve the problems of difficult observation, high false negative rate, observation that affects the results, etc., to reduce the occurrence of false positives and false negatives. Probability, accurate test results and high efficiency

Inactive Publication Date: 2006-07-05
CHONGQING FOOD IND RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is fast and simple, there are still some defects: one is that the yellow o-nitrobenzene easily diffuses into the agar after ONPG is decomposed, which affects the observation of the results; , is not easy to observe. Therefore, when Clark proposed to use this method to detect coliforms and E. coli in drinking water in 1990, there is a high false negative rate
In addition, there are some other methods for rapid detection of coliforms and E. coli, but many of them can only detect one kind of target bacteria in one experiment, that is, adding the enzyme substrate of one kind of bacteria to the culture medium to detect another kind of bacteria. One kind of target bacteria can only choose another medium, which not only increases the workload of the method detection, but also causes waste of raw materials and time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Preparation of medium for qualitative detection of coliforms and Escherichia coli.

[0018] (1) Weigh 3.5g of peptone, 2.0g of yeast extract, 0.6g of lactose, 0.05g of 4-methylumbelliferone-β-D-galactoside, 6.0g of sodium chloride, 2.5g of dipotassium hydrogen phosphate, Potassium dihydrogen phosphate 0.7g, sodium lauryl sulfate 0.1g, add distilled water to 1000mL and heat to dissolve;

[0019] (2) After the above solution is cooled, make up the solution to 1000mL with distilled water, adjust its pH value to 6.9±0.2, put it in an autoclave, heat it to 121°C±1°C, and keep it for 15-20 minutes to extinguish Bacteria, then, natural cooling;

[0020] (3) When the above solution is cooled to 50°C±5°C, add 25mL of indoxyl-β-D-glucoside solution with a concentration of 15mg / mL and 5.0mL of indoxyl-β-D-glucoside solution with a concentration of 1.1mg / mL of cefsulodin solution to obtain an agar-free medium for rapid qualitative detection of coliforms and E. coli in...

Embodiment 2

[0021] Example 2: Preparation of medium for qualitative detection of coliforms and Escherichia coli.

[0022] (1) Weigh 5.0g of peptone, 3.0g of yeast extract, 1.2g of lactose, 0.25g of 4-methylumbelliferone-β-D-galactoside, 7.5g of sodium chloride, 3.4g of dipotassium hydrogen phosphate, Potassium dihydrogen phosphate 1.2g, sodium lauryl sulfate 0.2g, add distilled water to 1000mL and heat to dissolve;

[0023] (2) After the above solution is cooled, make up the solution to 1000mL with distilled water, adjust its pH value to 6.9±0.2, put it in an autoclave, heat it to 121°C±1°C, and keep it for 15-20 minutes to extinguish Bacteria, then, natural cooling;

[0024] (3) When the above solution is cooled to 50°C±5°C, add 25mL of 32mg / mL indoxyl-β-D-glucoside solution and 4.5mL of 0.3mg / mL Cefsulodin solution, i.e. obtained agar-free medium for the rapid qualitative detection of coliforms and E. coli in drinking water, environmental water or food.

Embodiment 3

[0025]Example 3: Preparation of medium for qualitative detection of coliforms and Escherichia coli.

[0026] 1) Weigh 4g of peptone, 2.5g of yeast extract, 0.9g of lactose, 0.15g of 4-methylumbelliferone-β-D-galactoside, 6.5g of sodium chloride, 3g of dipotassium hydrogenphosphate, and 3g of dihydrogenphosphate Potassium 1g, sodium lauryl sulfate 0.15g, add distilled water to 1000mL and heat to dissolve;

[0027] (2) After the above solution is cooled, make up the solution to 1000mL with distilled water, adjust its pH value to 6.9±0.2, put it in an autoclave, heat it to 121°C±1°C, and keep it for 15-20 minutes to extinguish Bacteria, then, natural cooling;

[0028] (3) When the above solution is cooled to 50°C±5°C, add 20 mL of indoxyl-β-D-glucoside solution with a concentration of 25 mg / mL and 5 mL of indoxyl-β-D-glucoside solution with a concentration of 0.7 mg / mL that have just been prepared and sterilized by filtration. Cefsulodin solution, i.e. obtained agar-free medium...

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Abstract

This invention relates to a culture medium for the rapid detection of coliform and coliform groups, which is mixed from distilled water solution containing peptone, yeast extract, lactose, 4-methylumbelliferone-beta-D-galactoside, sodium chloride, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium lauryl sulfate and agar (or agar-free), the solution of indophenol-beta-D-glucoside and solution of cefsulodin. Peptone, yeast extract, lactose, 4-methylumbelliferone-beta-D-galactoside, sodium chloride, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium lauryl sulfate and agar (or agar-free) are weighed according to a certain criterion and added to distilled water for heat dissolution. The consequent solution is adjusted to a certain pH value, put into a high pressure sterilizer for sterilization and then added with filtrated and sterilized indophenol-beta-D-glucoside solution and cefsulodin solution after the temperature lowered to 50 deg.Cí‚5 deg.C. This is useful in either qualitative or quantitative detection of coliform and coliform groups in drinking water, environmental water and foods, high efficiency and accurate detection result.

Description

technical field [0001] The invention relates to sanitation and epidemic prevention detection and environmental water quality monitoring technology, in particular to a culture medium for rapid detection of coliforms and Escherichia coli in drinking water, environmental water or food and a preparation method thereof. Background technique [0002] In water or food, coliforms and Escherichia coli are used as the index bacteria for samples contaminated by enteropathogenic bacteria, and are one of the important indicators for evaluating water quality and food hygiene quality. Therefore, it can quickly and accurately detect water or Coliforms and Escherichia coli in food are of great significance. At present, there are two standards for the detection of coliforms and E. coli in food in China: one is the national standard, that is, the 9-tube fermentation method, which only stipulates the detection of coliforms. The detection steps are divided into three steps. The first step is th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04
Inventor 黄小平王瑜
Owner CHONGQING FOOD IND RES INST
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