A Duplex qPCR Assay Targeting the <i>fadA</i> Gene Enables Robust Detection of <i>Fusobacterium</i> in Clinical Samples.

ABSTRACT: Fusobacterium nucleatum (Fn) is increasingly recognized as a cancer-associated bacterium, yet reliable quantification in human specimens is challenging due to low bacterial burden and abundant host DNA. We analyzed 145 Fusobacterium genomes to design primers targeting conserved regions of the fadA adhesin gene and developed a duplex quantitative real-time PCR (qPCR) assay for simultaneous detection of fadA and a human PGT as an internal control. Analytical sensitivity, specificity, precision, and reproducibility were evaluated using serially diluted Fn DNA, spike-in experiments with human DNA, and cross-platform/operator validation. Clinical performance was assessed in colorectal cancer patient tissues, including fresh tissue (n = 24)

SUBMITTER: Seo Y 

PROVIDER: S-EPMC12691792 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature