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Fast detecting method for total number of bacterial colony in flavouring like sauce

A technology for the total number of colonies and detection methods, applied in biochemical equipment and methods, measurement/inspection of microorganisms, analysis through chemical reactions of materials, etc., to achieve rapid detection methods and low detection costs

Active Publication Date: 2009-01-14
FOSHAN HAITIAN FLAVOURING & FOOD CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The inventor believes that: for complex condiment samples such as soy sauce, the separation and enrichment of bacteria in the sample and the physiological state of the bacteria should be solved, and interfering substances should be removed; the bacteria should be repaired and rejuvenated to ensure the consi

Method used

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  • Fast detecting method for total number of bacterial colony in flavouring like sauce
  • Fast detecting method for total number of bacterial colony in flavouring like sauce

Examples

Experimental program
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Effect test

Embodiment 1

[0022] 1. Sample filtration Use a pipette to pipette 10ml of liquid food into 90ml of sterile saline, shake well, take 10ml of the diluted sample to pass through a 0.2μm-0.45μm membrane, collect the bacteria, and treat the bacteria trapped on the membrane Rinse three times with sterile normal saline to remove substances in condiments such as soy sauce that affect bacterial repair and interfere with luminescence. The data listed in Table 1 illustrate the presence of substances that interfere with luminescence in soy sauce.

[0023] Table 1 Effects on the luminescent system before and after removing interfering substances

[0024] sample name

relative luminescence value

(RLU, n=3)

Colony Count Results

(cfu / ml)

sample 1

(100μl light soy sauce, 10μl dilution solution + 100μl enzyme solution)

89507±1538

1700

sample 2

(100μl dark soy sauce, 10μl diluent + 100μl enzyme solution)

948±42

140

...

Embodiment 2

[0036] 1. To isolate the bacteria, use a balance to aseptically weigh 10g of solid or paste food into 90ml of sterile saline, shake it well, let it stand for 15min, take 20ml of the supernatant and centrifuge at 600g-1000g / 7min at 4°C to remove For food residues, take 10ml of the supernatant and pass it through a 0.2μm-0.45μm mixed cellulose ester microporous filter membrane, collect the bacteria, and wash the bacteria trapped on the membrane three times with sterile physiological saline to remove the bacteria that affect the repair in the food and substances that interfere with luminescence.

[0037] 2. Repair medium composition and preparation Medium formula: beef extract powder 2g, casein hydrolyzate 20g, soluble starch 0.5g, anhydrous glucose 7g, L-alanine 0.5g, inosine 0.5g, L-asparagine Vegetable 0.5g, add distilled water 1000ml. Sterilize at 121°C for 15 minutes and store at room temperature for use. The culture medium was divided into 180mm×18mm test tubes, 10ml per ...

Embodiment 3

[0041]1. To isolate the bacteria, use a balance to aseptically weigh 10g of solid or paste food into 90ml of sterile saline, shake it well, let it stand for 15min, take 20ml of the supernatant and centrifuge at 600g-1000g / 7min at 4°C to remove For food residues, take a 10ml sample of the supernatant and centrifuge at 6000g-10000g / 7min at 4°C, remove the supernatant, collect the bacteria, wash the bacteria three times with sterile saline repeatedly, and remove the substances in the food that affect bacterial repair and interfere with luminescence .

[0042] 2. Repair medium composition and preparation Medium formula: beef extract powder 5g, casein hydrolyzate 15g, soluble starch 3g, anhydrous glucose 3g, L-alanine 1g, inosine 0.25g, L-asparagine 2g , add 1000ml of distilled water. Sterilize at 121°C for 15 minutes and store at room temperature for use. The culture medium was divided into 180mm×18mm test tubes, 10ml per tube, and sterilized at 121°C for 15 minutes before use. ...

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Abstract

The invention discloses a rapid detecting method for detecting the total number of colonies in spices such as soy sauce. The method comprises the following steps: firstly, enriched thalli are separated by using the membrane separation technique, and substances affecting the growth and a luminescence system of the thalli are removed; secondly, a culture medium is used for repairing sublethal thalli and changing the spore into trophozoite cells, so the thalli are in sync growth status; and thirdly, a bacterial suspension and an ATP luminescence agent are reacted, so the total number of the colonies can be detected according to the light value. The invention discloses the culture medium used for detecting the total number of the colonies in spices quickly. The detecting method is rapid, the total number of colonies can be detected accurately within 4 hours, and the invention has the advantages of promptness and accuracy.

Description

technical field [0001] The invention relates to the technical field of condiments, in particular to a rapid detection method for the total number of bacterial colonies in condiments such as soy sauce. Background technique [0002] The total number of colonies in condiments such as soy sauce is an important indicator of food hygiene. At present, the detection method mainly uses the plate counting method, which requires 48 hours for detection, and there is often a hysteresis phenomenon. The improved method on this basis still needs to be cultivated for more than 12 hours, which cannot achieve the purpose of rapid detection. At present, it is reported in the literature that the ATP biological method can quickly detect the total number of colonies. This method uses the ATP of the bacteria itself as the detection object. However, after soy sauce and other condiments are pasteurized, most of the heat-labile bacteria are killed, and some are in a sublethal state. Some bacteria ha...

Claims

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Application Information

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IPC IPC(8): C12Q1/06G01N21/76
Inventor 严守雷缪素娜邹敏娟黄文彪潘思轶
Owner FOSHAN HAITIAN FLAVOURING & FOOD CO LTD
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