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Method for measuring intestinal tract setting capacity of lactobacillus

A determination method, the technology of Lactobacillus, applied in the field of microorganisms, can solve the problems of cumbersome operation, heavy workload, high cost, etc., and achieve the effects of high detection accuracy, fast operation, and reduced experiment cycle

Inactive Publication Date: 2015-11-11
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0019] The purpose of the present invention is to overcome the deficiencies of the prior art, overcome the limitations and time-consuming of the traditional plate culture method, the operation of the DGGE-PCR method is complex, the workload is large, the cost is high and the fluorescent labeling method is harmful to the tested object. Limitations and other defects provide a method for determining the intestinal colonization ability of Lactobacillus. Rapid and accurate quantification of six species of Lactobacillus Lactobacillus, Lactobacillus helveticus and Lactobacillus bulgaricus, and then compare their intestinal colonization ability

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  • Method for measuring intestinal tract setting capacity of lactobacillus
  • Method for measuring intestinal tract setting capacity of lactobacillus
  • Method for measuring intestinal tract setting capacity of lactobacillus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Example 1 Determination of Lactobacillus Intestinal Colonization Ability in Mice

[0100] SPF (Specific Pathogen Free) mice were used as the experimental model, and the colonization ability of the mice in the intestinal tract was determined by gavage of the mixed bacterial solution and later collection and detection of the amount of these six bacteria in the feces.

[0101] (1) Prepare six kinds of Lactobacillus bacteria powders of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus rhamnosus, Lactobacillus helveticus and Lactobacillus bulgaricus respectively, count the number of viable bacteria in the bacterial powder by plate counting, and prepare The mixed bacteria powder of six kinds of Lactobacillus ensures that the concentration of each bacteria is the same, and the mixed bacteria powder is stored at -20°C for later use.

[0102] (2) Purchase 4-6 week-old SPF mice and raise them in a barrier environment in the experimental animal c...

Embodiment 2

[0112]Example 2 Determination of Lactobacillus Colonization Ability in Human Intestines

[0113] Healthy adults were used as the experimental subjects, and the colonization ability of the strains in the human intestinal tract was determined by administering the mixed bacterial solution and later collecting and detecting the amount of these six bacteria in human feces.

[0114] (1) Prepare six lactobacillus powders, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus rhamnosus, Lactobacillus helveticus and Lactobacillus bulgaricus, and calculate the number of live bacteria in the powder by plate counting. Prepare a mixed bacterial powder containing six kinds of Lactobacillus to ensure that the concentration of each bacteria is the same, and store the mixed bacterial powder at -20°C for later use.

[0115] (2) 10 healthy adults aged 20-40 were organized to carry out the test, and the subjects' daily intake of 10 10 The mixed bacterial solution o...

Embodiment 3

[0123] The present invention provides six kinds of Lactobacillus, including Lactobacillus plantarum, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus rhamnosus, Lactobacillus helveticus, and Lactobacillus bulgaricus for measuring the intestinal colonization ability, specifically comprising the following steps:

[0124] (1) Collect the fresh feces of subjects who ingested a certain amount of mixed bacterial liquid / bacteria powder before and after ingesting the bacteria;

[0125] (2) Feces are quantitatively weighed to extract the bacterial genome in the feces;

[0126] (3) select the specific fluorescent quantitative PCR primers of these six lactobacilli of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus rhamnosus, Lactobacillus helveticus, Lactobacillus bulgaricus, and detect its amplification characteristics;

[0127] (4) respectively adopt the bacterial genome of six kinds of lactobacillus as template, adopt six pairs of corres...

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Abstract

The invention relates to a method for measuring the intestinal tract setting capacity of lactobacillus. The method comprises the following steps: 1, collecting excrement samples of a subject who intakes mixed bacterium liquid or bacterium powder; 2, extracting bacterium genomes in the excrement, and obtaining the excrement bacterium genomes; 3, selecting specific fluorogenic quantitative PCR primers, and detecting the amplification specificity of the primers; 4, adopting the bacterium genomes of six types of lactobacillus as templates respectively, and fabricating six types of standard curves between copy numbers of the six types of lactobacillus and ct values respectively; 5, conducting the fluorogenic quantitative PCR, and obtaining the ct values, wherein the amount of each type of lactobacillus in the unit excrement sample can be obtained through conversion; 6, comparing the lactobacillus content of excrement of the subject before and after the subject intakes the mixed bacterium liquid or the bacterium powder, and obtaining the setting capacity of bacterial strains in the intestinal tract. The method is based on the fluorogenic quantitative PCR, the rapid and accurate quantification of the six types of lactobacillus in the excrement samples can be achieved within a short time, and the intestinal tract setting capacity of the lactobacillus can be further compared.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a method for measuring the intestinal colonization ability of Lactobacillus. Intestinal colonization capacity of six species of Lactobacillus helveticus and Lactobacillus bulgaricus. Background technique [0002] Lactic acid bacteria is a general term for a group of bacteria that can use fermentable carbohydrates to produce a large amount of lactic acid. The more important genera of lactic acid bacteria are: Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, etc. Among them, Lactobacillus bacteria are widely used in food fermentation, industrial lactic acid production and medical care because of their good growth and fermentation characteristics and probiotic functions, and are closely related to human daily life. Lactobacillus has good fermentation characteristics and probiotic effects, which can not only improve the nutritional value of f...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/25C12R1/245C12R1/225
CPCC12Q1/6851C12Q1/689C12Q2531/113C12Q2537/16C12Q2563/107
Inventor 陈卫赵国忠王梦颖韩俊燕张灏赵建新田丰伟张秋香张白曦王刚刘小鸣郭敏
Owner JIANGNAN UNIV
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