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Method for rapidly detecting proteolysis bacteria in human excrement

A technology for protein hydrolysis and detection of human body, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., to achieve the effect of simple operation process, cost reduction, and easy operation

Inactive Publication Date: 2016-08-24
夏云
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, we have not seen reports on the use of in situ molecular microbial ecology methods to study the human intestinal microbial community (for example: the role of protein degrading bacteria in the utilization and transformation of nutrients, the present invention will provide metabolic diseases (diabetics) In situ information on the structure of protein-degrading bacterial communities in the gut microbiome and their nutritional functions

Method used

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  • Method for rapidly detecting proteolysis bacteria in human excrement
  • Method for rapidly detecting proteolysis bacteria in human excrement
  • Method for rapidly detecting proteolysis bacteria in human excrement

Examples

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Embodiment 1

[0045] A method for quickly detecting proteolytic bacteria in human feces, the process is as follows figure 1 As shown, including the following steps:

[0046] Step (1), collect total bacteria in feces under aseptic conditions:

[0047] (1.1) Sample collection: Use a sterile medicine spoon to randomly collect fresh human feces in a sterile bag, immediately store them in an incubator at 37°C, and send them to the laboratory for processing within 20 minutes;

[0048] (1.2) Dilute the sample: Take 10g of fresh stool, add it to 1×PBS buffer at pH 7.0, and mix well; the mass-volume ratio of the fresh stool to 1×PBS buffer W / V=0.95:3.5;

[0049] (1.3) Homogenization filtration: transfer the homogenized liquid in step (1.2) into a Bio-Rad homogenization bag, then place it in a Bio-Rad homogenizer and homogenize at 5000 R.PM for 8 minutes. After homogenization, The liquid in the homogenization bag is filtered through the filter grid on the side of the homogenization bag, and the filtrate is ...

Embodiment 2

[0060] A method for rapidly detecting proteolytic bacteria in human feces, including the following steps:

[0061] Step (1), collect total bacteria in feces under aseptic conditions:

[0062] (1.1) Sample collection: Use a sterile medicine spoon to randomly collect fresh human feces in a sterile bag, immediately store them in an incubator at 37°C, and send them to the laboratory for processing within 20 minutes;

[0063] (1.2) Dilute the sample: Take 20 g of fresh stool, add it to 1×PBS buffer at pH 8.0, and mix well; the mass-volume ratio of the fresh stool to 1×PBS buffer W / V=1.05:3.5;

[0064] (1.3) Homogenization filtration: transfer the homogenized liquid in step (1.2) into a Bio-Rad homogenization bag, and then place it in a Bio-Rad homogenizer to homogenize at 10000 R.PM for 15 min. After homogenization, Filter the liquid in the homogenization bag through the filter grid on the side of the homogenization bag, and take the filtrate;

[0065] (1.4) Centrifugation of the filtrate: ...

Embodiment 3

[0075] A method for rapidly detecting proteolytic bacteria in human feces, including the following steps:

[0076] Step (1), collect total bacteria in feces under aseptic conditions:

[0077] (1.1) Sample collection: Use a sterile medicine spoon to randomly collect fresh human feces in a sterile bag, immediately store them in an incubator at 37°C, and send them to the laboratory for processing within 20 minutes;

[0078] (1.2) Dilute the sample: Take 15g of fresh stool, add it to 1×PBS buffer at pH 7.5, and mix well; the mass-volume ratio of the fresh stool to 1×PBS buffer W / V=1:3.5;

[0079] (1.3) Homogenization filtration: transfer the homogenized liquid in step (1.2) to a Bio-Rad homogenization bag, and then place it in a Bio-Rad homogenizer to homogenize at 8000 R.PM for 10 minutes. After homogenization, The liquid in the homogenization bag is filtered through the filter grid on the side of the homogenization bag, and the filtrate is taken;

[0080] (1.4) Centrifugation of filtrate...

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Abstract

The invention relates to a method for rapidly detecting proteolysis bacteria in human excrement and belongs to the technical field of biological detection. The method completely adopts a culture-free molecular microbial ecology without separating strains and can conduct calibration and tracking on proteolysis bacteria in the human intestinal tract in an in-situ state. The method uses the intestinal tract proteolysis bacteria as a target serving as a long-term directional human indicator for health intervention and provides a data support for definition and diagnosis of the stability of human intestinal tract microbial functional flora. In addition, by means of detecting and calibrating the intestinal tract proteolysis bacteria, the relation between the physiological differences of individuals and the response to host energy metabolism can be found. Compared with existing pure culture methods, the method has the advantages that the method is rapid, low in cost and simple and convenient to operate, does not require special personnel training, has no special requirement for detection places, experimental results are accurate, and related data can be quickly provided for a diagnosis worker.

Description

Technical field [0001] The invention belongs to the technical field of biological detection, and specifically relates to a method for rapidly detecting proteolytic bacteria in human feces. Background technique [0002] The latest research shows that there are many kinds of bacteria in the human intestine. The intestine is another human “organ”. The composition and metabolism of microorganisms in the intestine are closely related to human health, such as obesity and diabetes (type II). The study of the composition and function of the complex human gut microbial community and how they affect the host's energy metabolism and the host's response mechanism to changes in the microbial community is an emerging frontier research field. The intestinal flora is a basic part of the human body, which has many effects on the nutrition, health and disease of the host. All studies have shown that the composition, abundance and function of intestinal microbes have important regulatory functions...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
CPCC12Q1/04
Inventor 夏云孔云虹黄鹤平王定康董宝财
Owner 夏云
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