In recent years, intestinal microbes can be said to be the hottest research hotspots, frequently on the cover of CNS journals, not only the academic papers on related topics are strong, but also the number of natural micro-projects sponsored by the country is also increasing. In 2018, nearly 180 studies were funded, with a total funding of 76 million . It can be seen that intestinal microbes have always been a research hotspot in the field of scientific research. It is believed that intestinal microbes in 2019 are still hotspots for national natural applications. In short, keeping up with this hot spot is not only easy to send a text, but also to send a good text, and it is easier to apply for a fund.
Insufficient existing research strategies
16S sequencing or macrogene is the most commonly used method for intestinal microbiology research, focusing on the correlation between intestinal microbial diversity and physiology and disease. Although such research methods can help to understand the flora migration in biological processes and identify potential strains involved in disease development, based on this, animal models can be used to use fecal transplantation or gavage experiments of specific strains. It can relatively directly reveal the causal relationship between intestinal microbes or specific strains and diseases, but little is known about the function and mechanism of the bacteria, which means that it is difficult to achieve the function of the flora by 16S sequencing or macro sequencing alone. Inquiry . Therefore, from the perspective of the integrity and innovation of the research proposal for the application of tenders by the country, this is a problem that needs everyone's attention.
The entry point for the study of intestinal flora function
At present, a large number of studies have found that intestinal microbes are functional and active for some small molecules produced during the metabolism of food or exogenous substances, such as SCFAs, bile acids, TMA/TMAO, neurotransmitters, amino acids and other substances on intestinal epithelial tissue. It also plays an important role in the metabolism of the whole body, immune regulation, and regulation of the central nervous system. The intestinal flora interacts with the host through these small active molecules . Therefore, in order to better understand the interaction between intestinal microbes and hosts, it is necessary not only to understand the diversity of intestinal flora, but also to further study the changes in intestinal flora function, ie, the intestinal flora metabolites. The changes were studied. Therefore, if the metabolites are used as a cutting point in the study of the function of the flora, it is believed that the integrity and innovation of the natural projects of the applicant country will be added a lot.
Open up the diversity + function research strategy - 16S + metabolome
The most common method for diversity analysis of the flora is to use the 16S rDNA amplicon sequencing method (hereinafter referred to as 16S), which is fast, efficient, and mature. The flora function can systematically detect the bacterial-related metabolites through metabolomics. Metabolomics is based on high-throughput analysis and bioinformatics technology, a omics technique for studying endogenous small molecules, which can objectively detect bacterial metabolites and clearly reflect the “function†of the intestinal flora. Changes under specific conditions. (If you don't pay attention to metabolites of special interest in the early stage, you can choose non-targeted metabolomics for large-scale screening. If you have metabolites of specific interest, you can choose to target metabolomics.) 16S combined with metabolomics, The combination of “diversity†and “function†of intestinal flora is the most biologically linked combination of two levels, which helps to fully explore the relationship between intestinal flora and disease occurrence, drug metabolism/drug efficacy.
It is precisely because 16S+ metabolism can be studied more deeply and completely, so many high-level articles in the past two years have adopted such research routines, and even many articles only supplemented a metabolic experiment, taking the data of the previous 16S again. Then published an article.
The key to the combined analysis of 16S+ metabolism - metabolomics
The 16S+ metabolic joint analysis can be done well, and the more critical ones may be in the metabolome experiment. 16S is based on sequencing technology. It started early and the method is mature. The detection capabilities and data quality of different platforms are relatively close. There is not much discussion here. Unlike 16S, metabolomics develops at the earliest and has certain specialities. It is far more complicated than transcriptomes and protein groups. For example, identification and quality control have many difficulties. The analytical capabilities of different platforms are still very different.
◠From the point of view of identification , the metabolome cannot be spliced ​​like sequencing, nor can it solve the identification problem directly through the public database like the proteome. Metabolites have a large number of isomers and substances of similar molecular mass, so if the retrieval of public databases by molecular mass tends to match multiple metabolites, such results are meaningless. How is the metabolite identified? When the instrument model and parameters are fixed, the secondary spectrum formed by the fragmentation of metabolites is fixed, but the metabolite interference matching results with similar fragmentation patterns can be avoided, so it is generally necessary to have experienced experimenters. Perform manual secondary confirmation. It is precisely because of metabolites have such characteristics, the current accurate identification of metabolites is very dependent on the local standard self-built database + manual secondary confirmation, is also an important basis for determining the reliability of the identification results of different analytical platforms.
â— From the perspective of quality control , the individualization of metabolism is large, so the number of samples is more than other group requirements. Therefore, experimental stability is a key point of investigation. It is also necessary to display quality control results . Similarly, different considerations are made. Quality control should also be a very important part of analyzing the platform, don't ignore it.
How to conduct a joint analysis of 16S+ metabolomics
Correlation analysis is a way of deep data mining, mainly used to discover the relevance or relevance hidden in the data set. The current 16S+ metabolomics joint analysis is divided into three steps:
The first step: the 16S and metabolome data were analyzed separately to screen for significant differential flora and significant differential metabolites.
Step 2: Calculate the correlation between significant differential flora and significant differential metabolites based on Spearman or Pearson statistical analysis. The greater the correlation coefficient, the stronger the correlation between the corresponding bacteria and metabolites.
The third step: by means of network diagrams, hierarchical clustering heat maps and other visualization methods, the correlation calculation results are displayed and excavated from multiple angles, and the key strains causing metabolic changes are screened.
to sum up
In summary, the 16S+ metabolic research strategy compensates for the lack of functional research on 16S or macrogenes. It is the most biologically significant two-group association, opening up the diversity and function of the flora, and relying on correlation analysis. The key strains and their functions can be mined, and the story of the article can be deeper, more complete and more innovative. The late development of metabolomics is the key to joint analysis. If there is a problem with the quality of metabolomics data, it will affect the joint analysis data mining.
Founded in 2004, Zhongke New Life (APT) is one of the earliest platforms for conducting metabolomics services in China. Compared with other platforms, APT has 2 advantages, which is worthy of your choice.
Special "new"
Zhongke New Life provides a complete and innovative solution:
â— Complete solution to diversity to function (16S+metabolism group).
â— Establish a macro proteomics service platform to provide a new perspective for the study of flora function.
To "quality"
Zhongke New Life has successfully built a sample of experienced, strict quality control and large project docking cooperation platform:
â— Experienced: The 2700+ metabolomics standard database has been successfully established, and the results are manually confirmed to ensure the reliability of the results.
â— Strict quality control: 6 quality control indicators are used to comprehensively assess the reliability of data quality.
â— Cooperation ability: It has published many high-level metabolomics articles, and participated in a number of precision medical special projects, fully capable of cooperating with large cohort projects.
Ducking! To the teachers who worked overtime on the weekend to write a national application for tenders!
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