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Hypervirulent Klebsiella pneumoniae (hvKp) is an emerging pathogen of increasing concern due to its ability to cause serious organ and life-threatening infections in healthy individuals and its increasing acquisition of antimicrobial resistance determinants. Identification of hvKp is critical for patient care and epidemiologic and research studies. Five genotypic markers on the hvKp-specific virulence plasmid can accurately differentiate hvKp from the less virulent classical K. pneumoniae (cKp) strain, but it is unclear whether the possession of fewer markers accurately predicts the hvKp pathotype. Likewise, the effect, if any, of various antimicrobial resistance factors on the pathogenic potential of hvKp has been incompletely explored. The Galleria mellonella infection model is often used to assess virulence, but this tool has not been validated. Therefore, levels of lethality of defined hvKp and cKp strain cohorts were compared in Galleria and outbred mouse models. The murine model, but not the G. mellonella model, accurately differentiated hvKp from cKp strains. Therefore, isolates in which the pathogenic potential is ambiguous due to an incomplete hvKp biomarker profile, an incomplete pLVPK-like hvKp-specific virulence plasmid, antimicrobial resistance that could decrease biofitness, and/or the lack of a characteristic clinical presentation should be validated in an outbred murine model. These data will assist in determining the minimal genomic content needed for full expression of the hypervirulence phenotype. This information, in turn, is critical for the development of the pragmatic point-of-care testing requisite for patient care and for the performance of epidemiologic and research studies going forward. IMPORTANCE Hypervirulent Klebsiella pneumoniae (hvKp) is of increasing concern because it can infect individuals in community and health care settings and because such infections are becoming difficult to treat. Identification of hvKp is important for patient care and to track its global spread. The genetic definition of hvKp, which can be used for its identification and the development of diagnostic tests, has not been optimized. Determination of possession of 4 of 5 genes that are present on the hvKp-specific virulence plasmid is highly accurate for identifying hvKp. However, an ongoing issue is whether strains that possess only some of these markers are still hypervirulent. The Galleria mellonella model and, less commonly, the murine infection model have been used to assess the virulence of these ambiguously identifiable strains. This report demonstrates that the murine model but not the G. mellonella model accurately identifies suspected hvKp strains. This information is critical for the development of diagnostics for patient care and for future research studies.

The Galleria mellonella Infection Model Does Not Accurately Differentiate between Hypervirulent and Classical Klebsiella pneumoniae