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Method for quickly detecting harmful bacteria in beer

A harmful bacteria, fast technology, applied in the field of beer, can solve the problems that the brewery cannot provide the contaminated bacteria, can not be qualitative, can not be distinguished, etc., to achieve the effect of convenient experimental observation and counting, intuitive observation, avoiding false positives and false negatives

Inactive Publication Date: 2013-02-06
CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this technology has the disadvantage of being indeterminate, so it cannot provide accurate information on the contaminating bacteria for the brewery. It is not known what kind of bacteria contaminates the beer, and it is impossible to distinguish whether it is a beer harmful bacteria.

Method used

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  • Method for quickly detecting harmful bacteria in beer
  • Method for quickly detecting harmful bacteria in beer
  • Method for quickly detecting harmful bacteria in beer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Take 1 bottle (600mL) of capped sake from a production workshop of a brewery, and after suction-filtering 100mL wine sample, take out the filter membrane and paste it on the rapid growth medium (peptone 8g, yeast extract 6g, beef extract 6g, spit temperature 800.8g, dipotassium hydrogen phosphate 2g, sodium acetate 6g, folic acid 0.2g, cycloheximide 6mg, agar 15g, glucose 15g, malt root extract powder 0.5g, calcium pantothenate 0.4g) plate, put After anaerobic culture in an anaerobic tank for 16 hours, perform fluorescent staining, place the polycarbonate membrane on sterile filter paper soaked in CFDA staining solution, and stain in the dark for 12 minutes, then take it out and place it in a sterile filter paper soaked in sterile water. Wash the staining solution under the membrane on the filter paper, then perform fluorescence in situ hybridization on the membrane according to the experimental procedure, add 10 μL volume of pre-designed Cy3-labeled Lactobacillus brevis...

Embodiment 2

[0030] Take 100mL of water from the production workshop of a brewery, filter the water sample, take out the filter membrane and paste it on the nutrient agar plate with the bright side up, and perform fluorescent staining after aerobic cultivation at 36°C for 5 hours, and put the polycarbonate membrane on the Wet the sterile filter paper soaked in CFDA staining solution, and stained in the dark for 12 minutes, then took it out and put it on the sterile filter paper soaked in sterile water to wash off the staining solution under the membrane, and then the membrane was fluorescent according to the experimental procedure In situ hybridization operation, wherein the fluorescent probe is a Cy3-labeled tetrad probe. After the treatment is completed, observe under the Olympus BX41 microscope, and record the experimental results.

[0031]

[0032] This method can quickly detect the number of microorganisms and the most common contamination of tetrad bacteria in beer.

[0033]

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Abstract

The invention discloses a method for quickly qualitatively and quantitatively detecting harmful bacteria in beer breweries by applying fluorescence in situ hybridization and fluorescent microcolony combined technology. The method comprises steps of: preparing sample solution and a culture medium; performing fluorescent microcolony and fluorescence in situ hybridization experimental operation; and observing by using a fluorescence microscope, counting, and qualitatively and quantitatively analyzing the harmful bacteria in the production process of the beer breweries according to fluorescent probes and excited fluorescence with different colors of a carboxyfluorescein diacetate (CFDA) dye. The invention also discloses two specific probes, which are respectively a probe for detecting Lactobacillus brevis, namely 5'-TAGCCGGTGTAACCCTGACTCTG-3' of which one end is marked by Cy3, and a probe for detecting micrococcus tetragenus, namely 5'-GAGATTAGGAAGAACACCAGTGGCGAA-3' of which one end is marked by Cy3. By combining two methods, phenomena that the fluorescence in situ hybridization is difficult to quantitate bacteria and experimental results are easy to show false negative and false positive are avoided, and the problem that the fluorescent microcolony technology cannot qualitatively analyze the bacteria is also solved. Therefore, the requirement on quick detection and qualitative and quantitative requirements for the production of the beer breweries are met.

Description

technical field [0001] The invention belongs to the technical field of beer. In particular, it relates to a method for rapid detection of harmful microorganisms in beer by applying the combination of fluorescence in situ hybridization technology and fluorescent micro-colony technology. Background technique [0002] Beer harmful bacteria refer to a type of bacteria that can survive anaerobically in packaged beer and cause beer to become sour and turbid. More than 85% of these bacteria are mainly composed of lactic acid bacteria, such as Lactobacillus brevis and Tetradococcus. The detection of harmful bacteria in beer production is a task that every brewery must face in order to avoid production loss, and it is also a major concern of the beverage industry around the world. In traditional beer microbial testing, the first step is often to screen microorganisms on selective media, which is a quantitative process. On the basis of the first step, further analysis is carried out...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
Inventor 王德良孟思严伟杰宋绪磊郭立芸穆英健沈建国
Owner CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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