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A method for detecting the relative content of bacterial species in a sample

A technology for relative content and detection of samples, applied in the biological field, can solve cumbersome problems and achieve the effect of overcoming cumbersome operations

Active Publication Date: 2021-02-09
深圳谱元科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] For this reason, the technical problem to be solved by the present invention is to simplify the cumbersomeness of the prior art, thereby proposing a method for detecting the relative content of the bacterial species in the sample

Method used

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  • A method for detecting the relative content of bacterial species in a sample
  • A method for detecting the relative content of bacterial species in a sample
  • A method for detecting the relative content of bacterial species in a sample

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Embodiment 1

[0052] Embodiment 1 This embodiment discloses a method for determining the relative content of Bifidobacterium and Lactobacillus in the total number of bacteria in a feces sample. The specific steps are as follows:

[0053] 1 sample pretreatment

[0054] 1.1 Spread a layer of plastic wrap on the surface of the ultra-clean workbench, and prepare weighing paper, disposable blades, and small spoons.

[0055] 1.2 Put on disposable PE gloves and take a half-thawed sample. Place the half-thawed sample on weighing paper, and cut off about 200 mg of the inner part (the part not exposed to the air) with a disposable blade before it is completely thawed, and put the cut part into a centrifuge tube with a blade or a spoon , complete the sampling.

[0056] 1.3 The sample after sampling is stored at -80°C. Throw the used weighing paper into the trash can, replace the disposable blade, put the blade into the sharps box, wipe and clean the sampling spoon with medical alcohol.

[0057] 1....

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a method for detecting the relative content of bacterial strains in a sample. The steps of the method are as follows: design of primers; genome extraction; fluorescent quantitative PCR; NGS16S sequencing; NGS analysis; Use NGS and qPCR to detect the relative content of a certain bacterium in the sample, and fit and correlate the NGS analysis results with the original qPCR experimental data to establish a predictive model, so that the subsequent qPCR analysis of the sample Test data can be input into the predictive model and directly obtain the relative content of a certain bacteria to be tested in the sample. The method reduces the complexity and cumbersome operation of the optimization of the relative content detection experiment of bacteria using qPCR alone, and improves the speed of obtaining the final result of the sample detection; it has great market prospects and economic value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for detecting the relative content of bacterial strains in a sample. Background technique [0002] As an important experimental technique, DNA sequencing has been widely used in biological research. In 1977, Sanger invented the landmark terminal termination sequencing method (generation sequencing). However, with the development of science, traditional Sanger sequencing can no longer meet the needs of research. The second-generation sequencing technology with lower cost, higher throughput and faster speed has emerged as the times require. The basic principle of next-generation sequencing is sequencing while synthesizing. On the basis of sequencing methods such as Sanger, through technological innovation, four different dNTPs (deoxy-ribonucleoside triphosphaTris-EDTA, deoxyribonucleoside triphosphates) are labeled with different colors of fluorescence. , the ne...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N1/28
CPCG01N1/28G01N21/6402G01N21/6486
Inventor 覃俊杰付志聪窦浩宇杨仁涛
Owner 深圳谱元科技有限公司
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