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.