Analysis method for diversity of oral cavity flora and disease-associated flora marker

An oral and sample technology, applied in the field of biotechnology detection, can solve problems such as deviation of research results

Inactive Publication Date: 2017-05-10
杨俊杰
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Problems solved by technology

However, due to the inevitable enrichment or attenuation of strains in traditional microbial culture techniques, the original micro-ecological structure is changed, which will lead to large deviations in the research results

Method used

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  • Analysis method for diversity of oral cavity flora and disease-associated flora marker
  • Analysis method for diversity of oral cavity flora and disease-associated flora marker
  • Analysis method for diversity of oral cavity flora and disease-associated flora marker

Examples

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Embodiment Construction

[0090] In Example 1 of the present invention, the samples to be tested are saliva samples from patients with systemic diseases, specifically, 32 samples of saliva samples from normal people, and 17 samples of saliva samples from patients with gastric cancer. A total of 49 disease samples were tested.

[0091] Using the above method, DNA was extracted from the oral flora of different patients, and the extracted DNA was subjected to PCR reaction, and the product of the PCR reaction was subjected to agarose gel electrophoresis, and then the PCR product was purified, and a library was constructed, ready to be used on the machine For sequencing, follow the steps below:

[0092] 1. Saliva sample sampling method

[0093] Sampling of saliva samples was carried out in strict accordance with the experimental procedures.

[0094] 2. Extraction method of saliva DNA

[0095] Salivary DNA was extracted by CTAB method.

[0096] 3. Determination of DNA content

[0097] The purity of the ...

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Abstract

The invention provides an analysis method for the diversity of oral cavity flora based on a second-generation high throughput sequencing technology and a disease-associated flora marker. The method comprises the following steps of (1) extracting a DNA of a to-be-detected saliva sample; (2) preparing a PCR reaction system, and carrying out PCR amplified reaction of V1-V2 regions of bacteria 16S rDNA, wherein the PCR reaction system comprises a PCR amplification template, a universal primer pair and the like; (3) carrying out detection of a target segment of a PCR product and purification of the PCR product; (4) carrying out second-generation high throughput sequencing on the purified PCR product and detecting the species of microorganisms in the saliva sample; (5) sequencing by using an Ion Torrent PGM sequencing system; and (6) carrying out bioinformatics analysis by using a FLASH, QIIME, usearch61. The diversity and the difference of the oral cavity flora of a patient are detected through the second-generation high throughput sequencing technology, so that the analysis method has the advantages that the composition and the functions of the flora can be more deeply analyzed in more detail by using the new-generation high throughput technology in comparison with a traditional flora detection method.

Description

technical field [0001] The invention relates to the field of biotechnology detection. Specifically, the invention provides a technology and method for detecting the diversity of oral flora in patients with diseases based on second-generation high-throughput sequencing technology. Background technique [0002] Hundreds of millions of microorganisms and their metabolites in the oral cavity play an important role in human energy metabolism, nutrient absorption, innate and acquired immunity, gastric and oral functions, etc. If it is broken, it will induce a variety of human diseases. At present, it has been recognized that oral microbes are closely related to human health and diseases. It is not enough to study human diseases only from the perspective of human beings. The role of oral microbes that live in symbiosis with humans must be considered. [0003] At present, there are many research methods applied to the diversity and differentiation of human oral microorganisms, main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6869C12Q1/689C12Q2531/113C12Q2535/122
Inventor 杨俊杰张磊姜梦菡
Owner 杨俊杰
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