Method for analyzing children autism intestinal flora diversity and specific primer pair

Abstract

The invention provides a method for analyzing children autism intestinal flora diversity and a specific primer pair. The method comprises the following steps: 1) extracting DNA of a to-be-detected stool sample; 2) configuring a PCR reaction system and performing PCR amplified reaction in bacteria 16S rDNA V1-V2 areas, wherein the PCR reaction system comprises a PCR amplification template, a universal primer pair and the like; 3) detecting a target fragment of a PCR product and purifying the PCR product; 4) performing second-generation high-throughput sequencing on the purified PCR product and detecting the types of microorganisms in the stool sample; 5) sequencing by utilizing an I11umina MiSeq sequencing system; 6) performing bio-information analysis by utilizing FLASH, QIIME and usearch61. The method provided by the invention has the benefits that through a second-generation high-throughput sequencing technology, the intestinal flora diversity of a diseased patient and the difference are detected; compared with a traditional flora detection method, the compositions and the functions of floras can be more detailedly and deeply analyzed by applying the new-generation high-throughput technology.

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 intestinal flora in patients with diseases based on next-generation high-throughput sequencing technology. Background technique [0002] Hundreds of millions of microorganisms and their metabolites in the intestinal tract play an important role in human energy metabolism, nutrient absorption, innate and acquired immunity, and gastrointestinal function. If the state is broken, it will induce a variety of human diseases. At present, it has been recognized that the gut microbes are closely related to human health and diseases. It is far from enough to study human diseases from the perspective of human beings alone. [0003] At present, there are many research methods applied to the diversity and differentiation of human intestinal microorganisms, mainly including traditional detection methods, gene lib...

AI Technical Summary

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

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