Biomedical subjects

Claudia Rotondo

Publications and source records attributed to Claudia Rotondo.

5 recordsLinked to original sources

PubMed2026

Parallel single-cell host immune profiling and pathogen genomic characterization in Klebsiella-associated sepsis: a pilot study.

OBJECTIVES: Sepsis is a life-threatening syndrome characterized by profound immune dysregulation and substantial biological heterogeneity. Here, we conducted a pilot study to explore host immune remodeling in Klebsiella-associated sepsis by combining single-cell RNA sequencing of peripheral blood mononuclear cells with whole-genome sequencing of the corresponding bloodstream isolates. METHODS: In this prospective observational pilot study, we analyzed peripheral blood mononuclear cells (PBMCs) from two patients with Klebsiella-associated sepsis and two healthy controls (HC) using single-cell RNA sequencing. PBMC composition, differential transcriptional responses, and pathway analysis were assessed across immune subsets. The corresponding bloodstream isolates were characterized by phenotypic antimicrobial susceptibility testing and whole-genome sequencing. RESULTS: Compared to HC, septic patients showed expansion of the myeloid compartment and contraction of the NK/T compartment. High-resolution analysis suggested shifts within lymphoid populations. At the transcriptional level, sepsis was associated with compartment-specific enrichment of interferon-related and host-defence pathways, as well as oxidative phosphorylation, ATP synthesis, and mitochondrial electron transport signatures across multiple PBMC subsets. Classical monocytes exhibited a coordinated decrease in MHC class II-related transcripts. The sepsis-associated isolates were identified as Klebsiella pneumoniae and Klebsiella variicola and were notable for overall antimicrobial susceptibility, limited resistomes, and absence of canonical hypervirulence determinants. CONCLUSION: Our data provide a preliminary description of immune remodeling during Klebsiella-associated sepsis and suggest that severe clinical disease may be associated with isolates lacking classical multidrug-resistance or hypervirulence features. These findings should be interpreted as preliminary and hypothesis-generating and require validation in larger cohorts with detailed clinical severity assessment.

Female↗

PubMed2026

A rapid molecular assay for the detection of hypervirulent Klebsiella pneumoniae in the context of antimicrobial resistance surveillance.

Hypervirulent Klebsiella pneumoniae (hvKP) represents an emerging clinical and public-health concern, particularly as hypervirulence increasingly converges with multidrug resistance. Current diagnostic approaches rely on phenotypic assays, such as the string test, or on whole-genome sequencing (WGS), both of which have limitations in specificity, turnaround time, standardization, and feasibility for routine surveillance. To address this gap, we developed a multiplex real-time PCR assay targeting key hvKP-associated virulence loci, including siderophore systems, hypermucoviscosity regulators, and additional markers linked to invasive potential. The assay was evaluated on 110 K. pneumoniae clinical isolates and 9 positive blood cultures, using WGS and the string test as comparators. The molecular panel demonstrated high concordance with WGS for principal virulence determinants, correctly identifying all high-virulence (score 4) profiles, and most intermediate profiles. Against WGS, the assay yielded a sensitivity of 82% and a specificity of 73%; performance against the string test was 96% and 87%, respectively. Direct testing from blood culture pellets yielded results consistent with both WGS and DNA-based PCR for the limited number of targets detected, supporting the technical feasibility of this approach. However, broader validation is needed to confirm performance in this specimen type. Overall, this multiplex PCR assay provides a targeted molecular screening approach for the rapid identification of hvKP-associated virulence profiles. Its agreement with genomic data supports its potential utility as an accessible complement to WGS for hvKP surveillance, although further workflow optimization will be required before broader routine implementation.IMPORTANCEThe global emergence of hypervirulent and multidrug-resistant K. pneumoniae represents a major public-health threat, as the convergence of virulence and antimicrobial resistance dramatically limits therapeutic options and increases the likelihood of severe, invasive, and potentially untreatable infections. Rapid identification of essential virulence determinants is therefore critical for timely clinical management and for preventing onward transmission. However, current diagnostic approaches are either insufficiently sensitive or require substantial resources, limiting their routine use. By providing a rapid and targeted molecular assay capable of detecting the principal loci associated with hypervirulent K. pneumoniae and by demonstrating the preliminary feasibility of its use directly on blood culture pellets previously identified as Klebsiella spp. by MALDI-TOF MS, this work provides a pragmatic approach for early virulence profiling. Implementation of such assays can significantly enhance epidemiological surveillance, support tailored patient management, and reduce the spread of high-risk K. pneumoniae lineages in both community and healthcare environments.

Klebsiella pneumoniae↗

PubMed2025

Rapid and accurate sepsis diagnostics via a novel probe-based multiplex real-time PCR system.

Sepsis is a critical clinical emergency that requires prompt diagnosis and intervention. Its prevalence has increased due to the aging population and increased antibiotic resistance. Early identification and the use of innovative technologies are crucial for improving patient outcomes. Modern methodologies are needed to minimize the turnaround time for diagnosis and improve outcomes. Rapid diagnostic tests and multiplex PCR are effective but have limitations in identifying a range of pathogens and target genes. Our study evaluated two novel probe-based multiplex real-time PCR systems: the SEPSI ID and SEPSI DR panels. These systems can quickly identify bacterial and fungal pathogens, alongside antibiotic resistance genes. The assays cover 29 microorganisms (gram-negative bacteria, gram-positive bacteria, yeast, and mold species), alongside 23 resistance genes and four virulence factors. A streamlined workflow uses 2 µL of broth from positive blood cultures (BCs) without nucleic acid extraction and provides results in approximately 1 h. We present the results from an evaluation of 228 BCs and 22 isolates previously characterized by whole-genome sequencing. In comparison to the reference methods, the SEPSI ID panel demonstrated a sensitivity of 96.88%, a specificity of 100%, and a PPV of 100%, whereas the SEPSI DR panel showed a sensitivity of 97.8%, a PPV of 89.7%, and a specificity of 96.7%. Both panels also identified additional pathogens and resistance-related targets not detected by conventional methods. This assay shows promise for rapidly and accurately diagnosing sepsis. Future studies should validate its performance in various clinical settings to enhance sepsis management and improve patient outcomes.IMPORTANCEWe present a new diagnostic method that enables the quick and precise identification of pathogens and resistance genes from positive blood cultures, eliminating the need for nucleic acid extraction. This technique can also be used on fresh pathogen cultures. It has the potential to greatly improve treatment protocols, leading to better patient outcomes, more responsible antibiotic use, and more efficient management of healthcare resources.

Humans↗

PubMed2006

Acute alcoholic hepatitis.

Acute alcoholic hepatitis (AAH) is a frequent inflammatory liver disease with high short-term mortality rate. In this review, relationships between alcohol abuse and the epidemiology and the outcomes of AAH are discussed, as well as AAH pathogenesis. The role of endotoxins, tumor necrosis factor alpha, fibroblasts, and immune response to altered hepatocyte proteins is discussed. The need of a careful prognosis, supported by the use of Maddrey score, by the model for end-stage liver disease [Mayo end-stage liver disease (MELD)] score or by the Glasgow alcoholic hepatitis score, is outlined, as the use of the most effective drugs (glucocorticoids and anti-tumor necrosis factor alpha infliximab) is recommended only in severe AAH cases. The problems of liver transplant in severe AAH, and the need of a 6-month alcohol abstinence before transplant, are discussed, as well as the need of a careful psychologic assessment before the transplant.

Acute Disease↗

PubMed2006

National Institute on Alcohol Abuse and Alcoholism and the study of fetal alcohol spectrum disorders. The International Consortium.

Fetal alcohol syndrome (FAS) is a large and rapidly increasing public health problem worldwide. Aside the full-blown FAS, multiple terms are used to describe the continuum of effects that result from prenatal exposure to alcohol, including the whole fetal alcohol spectrum disorders (FASD). The revised Institute of Medicine (IOM) Diagnostic Classification System and the diagnostic criteria for FAS and FASD are reported, as well as the formation of the four-state FAS International Consortium and its aims, as the development of an information base that systematizes data collection that helps to determine at-high-risk populations, and to implement and test a scientific-based prevention/intervention model for at risk women. The Consortium was further enlarged, with the inclusion of some more states (including Italy), leading to the formation of the International Consortium for the Investigation of FASD. The objectives of the Consortium are reported, as well as its previous activities, the South Africa and Italy Projects (active case ascertainment initiatives), and its future activities.

Adult↗