NGS data analysis method of enteric microorganism 16 SrRNA

A technology for gut microbes and data analysis, applied in the field of NGS data analysis of gut microbe 16SrRNA, can solve the problems of inaccurate data utilization in data analysis, and achieve the effect of improving data utilization, accurate results and cost saving

Pending Publication Date: 2021-03-02
申友基因组研究院(南京)有限公司
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Problems solved by technology

[0003] In order to solve the above problems, the present invention discloses an NGS data analysis method for 16S rRNA of intestinal microorganisms. The technical problem to be solved by the present invention is to realize the prob

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  • NGS data analysis method of enteric microorganism 16 SrRNA

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Embodiment

[0023] Embodiment: the method comprises the following steps:

[0024] 1) Database preparation: First, find out the sequence that can be amplified with the V4 universal primer from the SILVA132 and Dreengene13-8-16S databases. Two mismatches are allowed, and the V4 universal primer is cut off from the sequence to obtain a 254bp sequence. The 125bp sequences were intercepted in both directions to adapt to the actual situation of the sequencing sequence, and then the bidirectional 125bp sequences were connected to obtain a sequence with a length of 250bp;

[0025] 2) Feature classification trainer training: By using qiime2's naive Bayesian classifier to train the sequence in 1) and the classification information corresponding to the sequence and obtain a more accurate feature classifier, it can maximize the recognition of each classification group Sensitivity, specificity, precision and negative predictive value for most sequences in

[0026] 3) Accuracy assessment: use a simula...

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Abstract

The invention discloses an NGS data analysis method of enteromicroorganism 16 S rRNA, which comprises the following steps: 1), carrying out database preparation: finding out a sequence capable of being expanded by a V4 universal primer from SILVA132 and Duengene13-8-16S databases, cutting off the V4 universal primer from the sequence to obtain a 254bp sequence, respectively intercepting 125bp sequences from positive and negative directions, and connecting the bidirectional 125bp sequences; obtaining a sequence of which the length is 250bp; 2), training a feature classification trainer: training the sequence in the step 1) and the corresponding classification information by using a naive Bayes classifier of the qiime2 to obtain a more accurate feature classifier; 3), evaluating the accuracy, namely deducing the mixed sequencing sample by using a simulated microbial community and DADA2, and comparing the deduced mixed sequencing sample with real annotation information; and 4), performingdiversity analysis: executing multi-sequence alignment on the representative sequence by using a mafft program, and generating a phylogenetic tree based on the filtered alignment result by applying FastTree. The data utilization rate is improved, and the detection effect of longer read-length sequencing can be achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an NGS data analysis method of 16 S rRNA of intestinal microorganisms. Background technique [0002] The intestinal tract is not only a place for human digestion and absorption, but also the largest immune organ of the human body, which plays an extremely important role in maintaining the normal immune defense function of the human body. The human intestinal tract provides a good place for microorganisms to inhabit. The number of microorganisms in the adult intestinal tract is huge, weighing up to 1.2kg. In 2010, the EU Meta HIT project team published the gene catalog of human intestinal microbial army colonies in Nature. The genes contained in it The number is about 150 times that of the human body itself. It is estimated that there are at least 1000-1150 kinds of bacteria in the human intestinal tract. 16SrDNA is the most useful and commonly used molecular clock in the systematic...

Claims

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Application Information

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IPC IPC(8): G16B30/10
CPCG16B30/10
Inventor 董辉赵加栋金维荣秦红友周蓉
Owner 申友基因组研究院(南京)有限公司
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