Highly accurate and sensitive absolute quantification of bacterial strains in human fecal samples.

ABSTRACT:

Background

Next-generation sequencing (NGS) approaches have revolutionized gut microbiome research and can provide strain-level resolution, but these techniques have limitations in that they are only semi-quantitative, suffer from high detection limits, and generate data that is compositional. The present study aimed to systematically compare quantitative PCR (qPCR) and droplet digital PCR (ddPCR) for the absolute quantification of Limosilactobacillus reuteri strains in human fecal samples and to develop an optimized protocol for the absolute quantification of bacterial strains in fecal samples.

Results

Using strain-specific PCR primers for L. reuteri 17938, ddPCR showed slightly better reproducibility, but qPCR was almost as reproducible and showed comparable sensitivity (l

SUBMITTER: Li F 

PROVIDER: S-EPMC11380787 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature