3 results about "Bacterial 16S" patented technology

The application discloses a sequencing data calibration system and method based on microbial amplicon sequencing, and belongs to the field of microbial detection and bioinformatics analysis. The application takes ddPCR technology as the core, first screens common human intestinal microorganisms, designs and obtains 14 reconstructed sequences containing bacterial 16S and fungal 18S / ITS, prepares high-purity DNA standard substances through chemical synthesis and molecular cloning, completes standardization and uniformity verification, then designs high-specificity primers and probes, optimizes ddPCR reaction conditions, establishes a calibration method, completes uncertainty evaluation through joint calibration of 9 laboratories, and prepares multi-target amplicon abundance standard substances. The application realizes the transformation of amplicon sequencing data from relative quantification to absolute quantification, corrects the system error of the whole sequencing process, improves the comparability and reliability of cross-platform and cross-batch data, and provides technical support for the standardization of microbiome research.

The invention discloses a method for quality control of an internal standard substance of bacteria 16S rDNA and an absolute quantification method of bacterial flora, and relates to the technical field of molecular biology and sequencing. The specific primer and probe provided by the invention can simultaneously meet qPCR and dPCR platform detection, and can be used for quality control and period quality control of an internal standard substance and library quality control after a library is established by using the internal standard substance. The internal standard substance can realize relative quantification of bacteria 16S rDNA detection to absolute copy number quantification, and the precision and accuracy of 16S rDNA absolute quantitative sequencing are obviously improved. The quality control method provided by the invention has the obvious advantages of low quality control time cost and low economic cost. Besides, the invention also provides a method for absolute quantification of bacterial flora, and the method eliminates systematic deviation caused by DNA extraction efficiency and the like, and ensures that microbe abundance analysis in a 16S sequencing result is more accurate.

The present application relates to the technical field of molecular diagnosis, and specifically relates to application of female uterine cavity flora as a marker in endometriosis diagnosis. The present application collects uterine cavity flora of patients, extracts genomic DNA, performs bacterial 16s-rRNA sequencing, uses KEGG as a reference database for function prediction, determines that abnormal abundance of uterine cavity flora dominant bacteria has significant correlation with dysmenorrhea and high CA125 value of endometriosis patients, clarifies the correlation between abnormal uterine cavity flora and endometriosis, and provides a new method for evaluating the risk of endometriosis by detecting uterine cavity flora atlas.