Detection of methicillin ‐resistant Staphylococcus aureus persistence in osteoblasts using imaging flow cytometry

MRSA has been reported as the main pathogen involved in osteomyelitis and prosthetic joint infections. The host/pathogen interaction is dynamic and requires several changes to promote bacterial survival. Here, we focused on the internalization and persistence behavior ofStaphylococcus aureus invasive strains belonging to the major MRSA clones, within human MG ‐63 osteoblasts. We used imaging flow cytometry, a technique able to analyze living host cells. We demonstrated that the intracellular persistence process is different among clones and depends on the total number of infected cells instead of the number of intracellular bacteria. AbstractMethicillin ‐resistantS. aureus has been reported as the main pathogen involved in chronic infections, osteomyelitis, and prosthetic joint infections. The host/pathogen interaction is dynamic and requires several changes to promote bacterial survival. Here, we focused on the internalization and persistence behavior of well ‐characterizedStaphylococcus aureus invasive strains belonging to the main ST ‐MRSA‐SCCmec clones. To overcome the limitations of the cell culture method, we comparatively analyzed the ability of internalization within human MG ‐63 osteoblasts with imaging flow cytometry (IFC). After evaluation by cell culture assay, the MRSA clones in the study were all able to readily internalize at 3h postinfection, the persistence of intracellular bacteria was evaluated at 24h both by routine cell culture and IFC assay, ...

Source: MicrobiologyOpen - March 31, 2020 Category: Microbiology Authors: Dafne Bongiorno, Nicol ò Musso, Lorenzo Mattia Lazzaro, Gino Mongelli, Stefania Stefani, Floriana Campanile Tags: ORIGINAL ARTICLE Source Type: research