Qualitative and quantitative determination method of lactobacillus plantarum intestinal tract colonization

A kind of Lactobacillus plantarum, qualitative measurement technology, applied in the field of microorganism research, can solve problems such as high cost, heavy workload, complicated operation, etc., and achieves the effect of simple operation, high measurement efficiency, and simple qualitative measurement method.

Inactive Publication Date: 2012-10-03
BRIGHT DAIRY & FOOD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects of cumbersome operation, heavy workload and high cost in the existing denaturing gradient gel electrophoresis technique for determining DNA fragments of a specific sequence, and to provide a fast and simple method for the production of Lactobacillus plantarum Qualitative and quantitative determination methods of colonization

Method used

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  • Qualitative and quantitative determination method of lactobacillus plantarum intestinal tract colonization
  • Qualitative and quantitative determination method of lactobacillus plantarum intestinal tract colonization
  • Qualitative and quantitative determination method of lactobacillus plantarum intestinal tract colonization

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Embodiment 1

[0050] The qualitative assay method of the colonization of embodiment 1 lactobacillus plantarum

[0051] Contains 10 6 cfu / g Lactobacillus plantarum (lactobacillus plantarum) ST-III skimmed milk, Lactobacillus plantarum (lactobacillus plantarum) ST-III bacteria powder, Guangming Dairy Co., Ltd.

[0052] Tested fecal sample sampling: will contain 10 6 cfu / g Lactobacillus plantarum (lactobacillus plantarum) ST-III skimmed milk was refrigerated at 4-10°C, and the subjects ingested 100g of the skimmed milk daily for 3 days, and stopped taking the skimmed milk containing Lactobacillus plantarum From 3 days after milking, feces samples were taken once a day, and 0.1g-0.2g was aseptically collected.

[0053] The qualitative assay method of plantaractobacillus colonization comprises the steps:

[0054] (1) Extract the total DNA from the pretreated fecal samples;

[0055] The pretreatment is to freeze-dry the tested stool sample in liquid nitrogen, grind it, and use 10 g of sterile...

Embodiment 2

[0082] The qualitative assay method of the colonization of embodiment 2 lactobacillus plantarum

[0083] Lactobacillus plantarum (lactobacillus plantarum) 6003, Institute of Microbial Fermentation, Ministry of Light Industry; containing 10 8 cfu / g of Lactobacillus plantarum (lactobacillus plantarum) 6003 skimmed milk, Guangming Dairy Co., Ltd.

[0084] Tested fecal sample sampling: will contain 10 8 cfu / g Lactobacillus plantarum (lactobacillus plantarum) 6003 skimmed milk was refrigerated at 4-10°C, and the subjects ingested 100g of the skimmed milk daily for 3 days, and stopped taking the skimmed milk containing Lactobacillus plantarum From day 3, feces samples were taken once a day, 0.1g-0.2g were aseptically collected.

[0085] Measure according to the qualitative determination method of plantarum lactobacillus colonization in embodiment 1, determine whether each object has lactobacillus plantarum.

[0086] The experimental results of this embodiment are recorded in fi...

Embodiment 3

[0087] The qualitative assay method of the colonization of embodiment 3 lactobacillus plantarum

[0088] Lactobacillus plantarum (lactobacillus plantarum) ST-III, containing 10 8 cfu / g skimmed milk of Lactobacillus plantarum ST-III, Guangming Dairy Co., Ltd.

[0089] Tested fecal sample sampling: will contain 10 8 cfu / g Lactobacillus plantarum (lactobacillus plantarum) ST-III skimmed milk was refrigerated at 4-10°C, and the subjects ingested 100g of the skimmed milk daily for 3 days, and stopped taking the skimmed milk containing Lactobacillus plantarum From 3 days after milking, feces samples were taken once a day, and 0.1g-0.2g was aseptically collected.

[0090] Measure according to the qualitative determination method of plantarum lactobacillus colonization in embodiment 1, determine whether each object has lactobacillus plantarum.

[0091] The experimental results of this embodiment are recorded in image 3 in, such as image 3 as shown, image 3 It shows that the s...

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Abstract

The invention discloses a qualitative and quantitative determination method of lactobacillus plantarum intestinal tract colonization. The qualitative determination method comprises the following steps of (1) conducting total deoxyribonucleic acid (DNA) extraction on excrement samples to be measured and pretreated; (2) conducting polymerase chain reaction (PCR) amplification of total DNA; (3) conducting PCR amplification of a V3 area; (4) conducting denaturing gel gradient electrophoresis on products after the PCR amplification of the V3 area and conducting gel imaging gel analysis; and (5) enabling a stripe of denaturing gradient gel obtained in the step (4) to be compared with a blank group and a comparing marking group. The quantitative determination method comprises the following steps of operating samples to be measured and lactobacillus plantarum series concentration bacterium powder according to the step (1) to the step (4), then building a standard curve for luminance of a stripe on series concentration lactobacillus plantarum denaturing gradient gel and contents, and finally calculating colonization lactobacillus plantarum quantity according to the standard curve.

Description

technical field [0001] The invention belongs to the field of microbiology research, and relates to a qualitative and quantitative determination method for intestinal colonization of Lactobacillus plantarum. Background technique [0002] Lactobacillus plantarum is a type of Lactobacillus, most of which are isolated from plants. Lactobacillus plantarum inhibits the growth of pathogenic bacteria by competing with the ability of pathogenic bacteria to limit nutrients, regulates the composition of intestinal microecology, and forms a biological barrier. At the same time, through the effects of organic acids, bacteriocins, hydrogen peroxide and diacetyl produced by its metabolism on other bacteria, it can regulate the balance of intestinal microbial flora, enhance the body's immunity, reduce cholesterol levels, and inhibit tumor cells. formation etc. Lactobacillus plantarum has a variety of health-regulating and promoting functions in humans or animals. The prerequisite for thes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06C12R1/25
Inventor 徐致远刘振民艾连中于鹏
Owner BRIGHT DAIRY & FOOD
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