Genomic diversity of nontypeable H. influenzae strains confers phenotypic heterogeneity. Multiple strains of H. influenzae can be simultaneously isolated from clinical specimens, but we lack detailed information about polyclonal infection dynamics by this pathogen. A long-term barrier to our understanding of this host-pathogen interplay is the lack of genetic tools for strain engineering and differential labeling. Here, we present a novel plasmid toolkit named pTBH (
Keywords: Haemophilus influenzae; fluorescent labeling; mixed biofilms; mixed epithelial infection; plasmid toolbox; polyclonality assessment; selective antibiotic efficacy.