Development of a multiplex ddPCR assay for simultaneous absolute quantification of bacterial, fungal, and human DNA.

ABSTRACT: Molecular methods in clinical and research applications frequently encounter complex mixtures of human and microbial DNA, sometimes alongside environmental or reagent contaminants. In metagenomic studies, the presence of host contamination poses a significant challenge, reducing the assay sensitivity of microbial detection and characterization. Different host-depletion and microbial enrichment platforms have been developed to reduce or eliminate host DNA in samples predominantly composed of human DNA. To establish an effective method to assess the efficacy of host DNA depletion or microbial enrichment platforms, this study aimed to develop a multiplex, broad-range 16S/18S ribosomal DNA droplet digital PCR (rDNA ddPCR) assay capable of simultaneously quantifying bacterial and fungal DNA, al

SUBMITTER: Huang G 

PROVIDER: S-EPMC12923063 | biostudies-literature | 2026

REPOSITORIES: biostudies-literature