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Reagent kit and testing method for rapidly evaluating whether intestinal microecology of human body is unbalanced

A technology of intestinal microecology and kits, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, fluorescence/phosphorescence, etc., and can solve problems such as the DNA yield of bifidobacteria and enterobacteria

Inactive Publication Date: 2013-04-17
ZHEJIANG UNIV
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Problems solved by technology

However, the absolute quantification method used involves the preparation of Bifidobacterium and Enterobacter standard products and the establishment of a standard curve, and does not consider the yield of Bifidobacterium and Enterobacter DNA in stool samples

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  • Reagent kit and testing method for rapidly evaluating whether intestinal microecology of human body is unbalanced
  • Reagent kit and testing method for rapidly evaluating whether intestinal microecology of human body is unbalanced
  • Reagent kit and testing method for rapidly evaluating whether intestinal microecology of human body is unbalanced

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Embodiment Construction

[0082] Table 1 shows the quantitative detection results of the six dominant bacteria in the intestinal tract of 32 healthy young and middle-aged subjects, 31 patients with chronic hepatitis B, and 31 patients with decompensated liver cirrhosis. The results showed that the intestinal microecology of 31 patients with chronic hepatitis B and 31 patients with decompensated liver cirrhosis were disturbed, and the intestinal microecology of 31 patients with decompensated liver cirrhosis was seriously unbalanced.

[0083] Table 1

[0084]

[0085] Note a The data is the mean ± variance, and the unit is the copy number of the specific gene of the bacteria to be tested contained in each gram of stool (wet weight); the detection limit is

[0086] 5*10 3 Copy number / gram of stool (wet weight); *P<0.01, compared with the healthy control group, the difference is significant; ΔP<0.01, compared with the chronic hepatitis B patient group, the difference is significant;

[0087] Table 2 ...

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Abstract

The invention relates to the field of microbial ecology, and aims at providing a reagent kit and a testing method for rapidly evaluating whether intestinal microecology of a human body is unbalanced. The reagent kit detects a level ratio of bifidobactirium to enteric bacilli in feces by a relative fluorescence quantitative PCR (polymerase chain reaction) technique, and then evaluates the unbalance degree of the intestinal microecology. The method comprises the steps that a bifidobactirium and enteric bacilli quantitative detection primer with best specificity and high amplification efficiency is selected; the DNA yield (deoxyribose nucleic acid) of bifidobactirium and enteric bacilli is calculated; microorganisms are subjected to total DNA extraction; relative quantitative detection of bifidobactirium DNA and enteric bacilli DNA in total DNA is conducted; an X value is calculated through a formula; and the unbalance degree of the intestinal microecology is evaluated according to the X value. The reagent kit optimizes bifidobactirium and enteric bacilli quantitative detection conditions, has the advantages of high speed, good accuracy and high sensitivity, can be used for clinically evaluating a therapeutic effect of a microecological modifier, and can also be used for guiding administration of drugs for clinical digestive system dysfunction.

Description

technical field [0001] The invention relates to a rapid assessment kit for human intestinal microecological imbalance, which belongs to the field of microbial ecology. Background technique [0002] The number of microorganisms that colonize the human gut is about 10 14 1, which is equivalent to 10 times the number of human own cells, and the total number of genes they carry is 100 times the total number of genes carried by the human body. Gut microbes are recognized as an important organ of the human body because they are responsible for the metabolism of nutrients and immune functions of the body. These microorganisms are combined in a certain proportion, and through the regulation mechanism of the host and itself, a dynamic and relatively balanced state is achieved in the intestinal tract of the host, which plays an important role in the health of the human body. It has been recognized that changes in the structure and function of intestinal microecology are related to h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10C12Q1/06G01N21/64
Inventor 李兰娟鲁海峰吴仲文杨介钻张华陈燕飞
Owner ZHEJIANG UNIV
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