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Primer compound for identifying intestinal microecological state and application thereof

A technology of primer composition and intestinal microecology, which is applied in the direction of microorganisms, methods based on microorganisms, measurement/inspection of microorganisms, etc., can solve the problems of unpredictability of the overall situation of intestinal microecology and strong limitations, and achieve improved detection Specificity and sensitivity, the effect of improving detection specificity

Active Publication Date: 2019-04-05
SUZHOU PRECISION BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has strong limitations and cannot predict the overall situation of intestinal microecology

Method used

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  • Primer compound for identifying intestinal microecological state and application thereof
  • Primer compound for identifying intestinal microecological state and application thereof
  • Primer compound for identifying intestinal microecological state and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] The assembly of embodiment 1 kit

[0127] The design of primer composition, concrete sequence is as shown in table 1:

[0128] Table 1

[0129]

[0130]

[0131]

[0132] PCR reaction solution: Tris-HCl: 10-80mmol / L, pH8.5; Ammonium sulfate: 10-40mmol / L; MgCl 2 : 1-5.5mmol / L; dNTPs: 100-500μmol / L; final primer concentration: 0.25-1.25ng / μL. The components of the reaction solution in each reaction well are as follows:

[0133] Clostridium symbiotic PCR reaction solution: Tris-HCl: 10mmol / L, pH8.5; Ammonium sulfate: 10mmol / L; MgCl 2 : 3mmol / L; dNTPs: 200μmol / L;

[0134] Bifidobacterium adolescentis PCR reaction solution: Tris-HCl: 40mmol / L, pH8.5; Ammonium sulfate: 30mmol / L; MgCl 2 : 2.5mmol / L; dNTPs: 150μmol / L;

[0135] Fusobacterium nucleatum PCR reaction solution: Tris-HCl: 60mmol / L, pH8.5; Ammonium sulfate: 40mmol / L; MgCl 2 : 5.5mmol / L; dNTPs: 400μmol / L;

[0136] Anaerobic Peptostreptococcus PCR reaction solution: Tris-HCl: 80mmol / L, pH8.5; Ammonium s...

Embodiment 2

[0148] Embodiment 2 clinical verification

[0149] 1. Sample selection

[0150] 46 samples of intestinal microecological imbalance were collected in the hospital. All samples were confirmed by stool microscopy and doctors, including 8 cases of intestinal polyps and 38 cases of adenoma. At the same time, 28 healthy people with healthy physical examination were collected. The physical examination indicators were normal, and the fecal microscopy examination was normal. All of them had undergone colonoscopy examination and had no intestinal diseases.

[0151] 2. Genome extraction from stool samples

[0152] The bacterial genomes in different samples were extracted according to the instructions of the Fecal Bacterial Genome Extraction Kit (DP328, Tiangen Biochemical Technology (Beijing) Co., Ltd.), and the concentration and quality of DNA were detected.

[0153] 3. Check the samples to be tested

[0154] The extracted DNA was used as a template, and the loading amount was 5ng / sa...

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Abstract

The invention provides a primer compound for identifying intestinal microecological state and an application thereof. The primer compound is a specific primer designed for intestinal bacterium 16S rRNA; the specific primer includes random basic groups; the quantity of random basic groups is 3-5; the intestinal bacterium includes any one of or combination of at least two of symbiotic clostridia, bifidobacterium adolescentis, fusobacterium nucleatum, peptostreptococcus anaerobius or klebsiella. A kit researched and developed on the basis of the primer compound provided by the invention has a specificity for detecting intestinal microecological imbalance reaching up to 92.5% and a sensitivity reaching up to 80.43%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a primer composition and its application, mainly to a primer composition for identifying the intestinal microecological state and its application, in particular to a primer composition for identifying the intestinal microecological state and kits prepared therefrom. Background technique [0002] In recent years, the incidence of intestinal diseases has been rising, and the early symptoms of intestinal diseases are not obvious, and there is a lack of effective screening methods, which may eventually develop into cancer. Examination and screening of intestinal diseases in the prior art mainly screen high-risk groups through fecal occult blood test, followed by colonoscopy. However, the fecal occult blood test lacks specificity, the rate of colonoscopy examination for primary screening compliance and high-risk groups is only 30-40%, and the disease detection rate is about 19%, which need...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/10C12Q1/04C12N15/11C12R1/145C12R1/01C12R1/46C12R1/22
CPCC12Q1/689
Inventor 朱永亮穆延召杨敏靖星宇张水龙王一伟
Owner SUZHOU PRECISION BIOTECH CO LTD
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