Isolation and Characterization of Fusobacterium nucleatum Subspecies from Oral Clinical Specimens

The identification and quantification of Fusobacterium nucleatum subspecies is of paramount importance to researchers investigating fusobacterial composition in clinical specimens of human health and disease. Here, we present two validated methods to identify and/or quantify oral fusobacteria present within clinical specimens. Importantly, both approaches can unambiguously distinguish the four F. nucleatum subspecies and Fusobacterium periodonticum. The first method is a convenient PCR-based approach used for rapid genotyping of individual fusobacterial colonies and mixed cultures. The second is a quantitative, next-generation sequencing-based approach that directly measures the different fusobacteria present within complex clinical specimens. Together, these methods provide a comprehensive toolkit for fusobacterial profiling in clinical studies.