Early and high Influenza A circulation alongside reduced Respiratory Syncytial Virus prevalence during autumn 2025 in paediatric patients attending emergency department Velia Chiara Di Maio, Rossana Scutari, Stefania Ranno, Luana Coltella, Venere Cortazzo, Luna Colagrossi, Anna Chiara Vittucci, Livia Antilici, Giulia Linardos, Alessandra Salvatori, Leonarda Gentile, Manuela Onori, Barbara Lucignano, Gianluca Vrenna, Mara Pisani, Sebastian Cristaldi, Massimiliano Raponi, Cristina Russo, Paola Bernaschi, Alberto Villani, Carlo Federico Perno Italian Journal of Pediatrics, 2026 The recent implementation of preventive strategies against Respiratory Syncytial Virus (RSV) in infants has raised questions about potential changes in RSV seasonality and the circulation of other respiratory viruses. This study aimed to compare respiratory virus activity among paediatric patients presenting to the Bambino Gesù Children's Hospital (Rome, Italy), with respiratory symptoms and available BioFire Respiratory Panel 2.1 plus results during September-December 2024 (N = 670) and 2025 (N = 905). RSV detection decreased significantly in December 2025 compared with the same period in 2024 (6.3% vs 29.4%, risk ratio [RR]: 0.21, 95% confidence interval [CI] 0.16-0.28; p < 0.001). By contrast, influenza A viruses had an earlier onset in 2025 and emerged as the dominant pathogen compared with 2024 (21.0%, in 2025 vs 4.5%, in 2024; RR: 4.69, 95% CI 3.23-6.80; p < 0.001). Unlike September-December 2024, parainfluenza viruses (PIVs) 3 and 4 circulated in 2025 (0.4% vs 3.5% for PIV 3; 0.0% vs 3.6% for PIV-4; p < 0.001 for both), mainly observed in infants (median age 0.7 years; interquartile range: 0.2-1.2; mean±standard deviation: 0.9 ± 0.8). Circulation of adenovirus (11.5%, vs 6.1%; p < 0.001) and human coronavirus HKU1 (3.6%, vs 0.1%; p < 0.001) also increased in 2025 compared with 2024. The observed epidemiological shifts suggest potential effects associated with the introduction of RSV prophylaxis in Italy and highlight the importance of continued paediatric surveillance in monitoring evolving viral patterns and guiding public health decisions.
Furuncular myiasis by Cordylobia anthropophaga in three Italian children: First case series and implications for non-endemic regions Livia Mancinelli, Marika Guercio, Gianluca Vrenna, Manuela Onori, Claudio De Liberato, Adele Magliano, Andrea Diociaiuti, Massimiliano Raponi, Carlo Federico Perno, May El Hachem, Paola Bernaschi Plos Neglected Tropical Diseases, 2026 Cordylobia anthropophaga, commonly known as the tumbu fly, is a leading cause of cutaneous myiasis in sub-Saharan Africa. This condition is characterized by a papulopustular lesion that evolves in a painful boil-like nodule with central ulceration. Human infestations typically occur on covered body parts, due to the fly’s habit of laying eggs on damp clothing. This report describes the first cluster of three pediatric cases of furuncular myiasis caused by C. anthropophaga , diagnosed at the Bambino Gesù Children’s Hospital (Rome, Italy) between April 2024 and April 2025. All patients had recent travel history to endemic African regions and presented with cutaneous nodules, some accompanied by systemic signs such as fever or regional lymphadenopathy. Diagnosis was confirmed through clinical evaluation and morphological identification of the extracted larvae. Treatment consisted of occlusive therapy to facilitate larval expulsion or surgical extraction of the maggots and systemic antibiotics. One patient also exhibited a family cluster, with larval extraction from a parent. Laboratory results were largely unremarkable, consistent with localized infection. These cases highlight the growing relevance of imported myiasis in non-endemic countries like Italy, due to increased international travel and potential climate-driven changes in vector distribution. Enhanced awareness among travelers and healthcare providers, together with proactive public health measures, is crucial. By documenting these cases, we hope to contribute to the understanding of emerging parasitic diseases in non-endemic regions and reinforce the need for vigilance in this time of global environmental changes.
Expanding Diagnostic Options for Pediatric Meningitis: BCID2 Testing Results on Cerebrospinal Fluid After a Negative Meningitis/Encephalitis Panel Venere Cortazzo, Lorenza Romani, Gianluca Vrenna, Maia De Luca, Marilena Agosta, Martina Rossitto, Valeria Fox, Barbara Lucignano, Manuela Onori, Stefania Mercadante, Vito Tommaso, Laura Lancella, Stefania Bernardi, Mara Pisani, Alessandra Salvatori, Alberto Villani, Massimiliano Raponi, Carlo Federico Perno, Paola Bernaschi Antibiotics, 2026 Background: Rapid etiological diagnosis of bacterial meningitis is crucial in children, as delays can lead to neurological sequelae. The BioFire FilmArray Meningitis/Encephalitis (ME) panel is widely used on cerebrospinal fluid (CSF), but its target spectrum may miss healthcare-associated or multidrug-resistant pathogens. We evaluated the diagnostic performance and stewardship-oriented clinical impact of off-label BioFire FilmArray Blood Culture Identification 2 (BCID2) testing on CSF from pediatric patients with suspected bacterial CNS infection and negative ME results. Methods: We retrospectively analyzed CSF samples collected between January 2023 and March 2025 at a tertiary pediatric hospital. In ME-negative cases with persistent suspicion and abnormal CSF parameters, BCID2 was performed off-label on residual CSF aliquots after routine testing, without additional sampling. We assessed pathogen detection, agreement with culture, resistance-gene identification, and documented stewardship actions. Results: Among 76 ME-negative CSF samples tested with BCID2, 23 (30.3%) were positive, all involving organisms not included in the ME panel. BCID2 was concordant with culture in 19/23 cases (82.6%); 4/23 (17.4%) were BCID2-positive/culture-negative, consistent with reduced culture sensitivity in frequently pretreated cases. Resistance genes (VIM, vanA/B, CTX-M) were detected in 30.4% of BCID2-positive samples. Overall agreement with culture was 94.7% (PPA 100%, NPA 93.0%). Escalation was documented in 13/23 episodes (56.5%), discontinuation in 2/23 (8.7%), and confirmation in 9/23 (39.1%), with no de-escalation events; clinical outcomes were not systematically available. Conclusions: In selected ME-negative pediatric cases with abnormal CSF profiles, BCID2 testing on residual CSF provided rapid, clinically meaningful microbiological information that may support antimicrobial optimization.
Machine Learning Model with Fourier-Transform Infrared Spectroscopy (FTIR) as a Proof-of-Concept Tool for Predicting Group A Streptococcus (GAS) emm-Type in the Pediatric Population Valeria Fox, Gianluca Vrenna, Martina Rossitto, Serena Raimondi, Marco Cristiano, Venere Cortazzo, Marilena Agosta, Barbara Lucignano, Manuela Onori, Vanessa Tuccio Guarna Assanti, Maria Stefania Lepanto, Nour Essa, Isabella Tarissi De Jacobis, Andrea Campana, Massimiliano Raponi, Alberto Villani, Carlo Federico Perno, Paola Bernaschi Diagnostics, 2025 Background: Since 2022, invasive Group A Streptococcus (GAS) infections have increased, mainly due to the spread of specific emm-types, such as emm1. As therapy may depend on the emm-type, rapid and cost-effective identification is crucial. Fourier-transform infrared spectroscopy (FTIR) has emerged as a promising alternative to sequencing for GAS typing. We applied machine learning (ML) to FTIR spectra to build a predictive model for emm-type identification. Methods: Twenty-four GAS strains were analyzed by whole-genome sequencing and FTIR. The model was trained on twenty-one strains (emm-types: 1, 3, 4, and 6), using leave-one-out cross validation (LOOCV). To test the model’s ability to avoid false positive results, the model was also tested with three strains belonging to emm-types not included in the training of the model (emm-types: 12, 89, and 75). Results: An artificial neural network trained for 400 cycles achieved the highest accuracy (90.7%) out of the thirteen different models tested. When the three strains belonging to emm-types not included in the model were predicted with this model, it produced low score values, confirming its ability to avoid false positive results. Conclusions: We developed a preliminary and proof-of-concept model capable of accurately predicting the four most-prevalent emm-types in our setting, including the highly virulent emm1. These findings support FTIR combined with ML as a rapid, low-cost tool for GAS typing, with potential for real-time clinical applications to guide timely treatment decisions. However, as a proof-of-concept study, the relatively small sample size and limited emm-type diversity underline the need for further validation with larger and more diverse datasets.
When Conventional Methods Fail: First Detection of a Candida viswanathii Outbreak in Europe in a Pediatric Hospital Revealed by Whole Genome Sequencing and FT-IR Spectroscopy Gianluca Vrenna, Valeria Fox, Venere Cortazzo, Serena Raimondi, Marco Cristiano, Gianluca Foglietta, Sara Carilli, Martina Rossitto, Barbara Lucignano, Manuela Onori, Maria Paola Ronchetti, Andrea Dotta, Andrea Campana, Lorenzo Galletti, Luca Di Chiara, Alberto Villani, Marta Luisa Ciofi Degli Atti, Daniela Perrotta, Corrado Cecchetti, Massimiliano Raponi, Carlo Federico Perno, Paola Bernaschi Microorganisms, 2025 Candida viswanathii has been sporadically reported in Asia and South America but not in Europe. This study reports the first European outbreak of C. viswanathii in a paediatric hospital, outlining diagnostic challenges and containment measures. Fifteen C. viswanathii isolates were recovered from blood cultures of consecutive pediatric patients admitted to intensive care units between April and August 2025. Identification was performed using MALDI-TOF MS, chromogenic media, Fourier-transform infrared (FT-IR) spectroscopy, and whole-genome sequencing (WGS). Antifungal susceptibility testing was performed by broth microdilution. All isolates were initially misidentified as C. tropicalis by MALDI-TOF MS and undetected by the FilmArray BCID2 panel. WGS confirmed C. viswanathii, and FT-IR analysis revealed clonally related strains, indicating an outbreak. Colonies displayed a distinct deep-blue color on chromogenic CHROMagar™ medium. Elevated fluconazole minimum inhibitory concentrations were observed, while isolates remained susceptible to echinocandins and amphotericin B. A multidisciplinary infection-control response halted transmission within four weeks. This investigation documents the first C. viswanathii outbreak in Europe, highlighting diagnostic limitations of current commercial tools and the need for updated databases. Integration of FT-IR spectroscopy and WGS facilitated outbreak detection and containment, underscoring the importance of advanced diagnostics and surveillance for emerging fungal pathogens.
Unraveling Pediatric Group A Streptococcus Meningitis: Lessons from Two Case Reports and a Systematic Review Lavinia Di Meglio, Maia De Luca, Laura Cursi, Lorenza Romani, Mara Pisani, Anna Maria Musolino, Stefania Mercadante, Venere Cortazzo, Gianluca Vrenna, Paola Bernaschi, Roberto Bianchi, Laura Lancella Microorganisms, 2025 Streptococcus pyogenes meningitis is a rare invasive disease, accounting for less than 2% of bacterial meningitis. We presented two case reports and conducted a systematic review using PUBMED, covering the database from its inception up to 31 December 2024, of pediatric cases of Streptococcus pyogenes meningitis. Only case reports and case series were included. Differences in clinical and laboratory parameters were compared between uneventful course and complicated admissions. A total of 57 cases were included. The median age at diagnosis was 4 years. A primary infection focus outside the brain was identified in 61.39% of cases. S. pyogenes was identified from cerebrospinal fluid in 66.66% of cases and from blood in 15.79%. Septic shock occurred in 24.56% of cases, and 36.84% had brain anatomical anomalies. All patients received broad-spectrum empiric antibiotics, while protein-synthesis inhibitors were administered in 26.31% of cases. A total of 17% of patients died, and 28.07% experienced sequelae. The identification of S. pyogenes from blood and a Phoenix Sepsis Score ≥ 2 were significantly associated with a complicated clinical course. Our findings may offer useful insights for the clinical management of Streptococcus pyogenes meningitis.
An Evaluation of a Syndromic Molecular Panel in Optimising the Microbiological Diagnosis and Antimicrobial Therapy of Suspected Osteoarticular Infections in Paediatric Patients Marilena Agosta, Venere Cortazzo, Manuela Onori, Barbara Lucignano, Gianluca Vrenna, Martina Rossitto, Maria del Carmen Pereyra Boza, Valeria Fox, Marco Roversi, Antonio Musolino, Andrzej Krzysztofiak, Laura Lancella, Marco Giordano, Francesco Falciglia, Ottavia Porzio, Alberto Villani, Carlo Federico Perno, Paola Bernaschi Diagnostics, 2025 Background/Objectives: Paediatric osteoarticular infections (POAIs) present unique diagnostic and therapeutic challenges. Microbiological culture (MC) is typically time-consuming and lacks sensitivity, especially when patients have received antibiotics. The BIOFIRE® Joint Infection Panel (BJIP) is a syndromic molecular assay for the direct identification of most pathogens causing POAIs. Methods: We evaluated BJIP in 17 synovial fluids, and then, we retrospectively assessed its utility in 93 off-label specimens (i.e., 25 purulent fluids/biopsies and 68 whole blood samples). All specimens were collected from October 2022 to March 2024 from paediatric patients admitted at the Bambino Gesù Children’s Hospital in Rome. Results: A bacterial pathogen was isolated in only one of 17 synovial fluid cultures, while BJIP identified eight additional microorganisms in MC-negative cases. The most frequently detected pathogen was S. aureus (44.5%, 4/9). BJIP performance in synovial fluids showed an overall positive percentage agreement (PPA) and negative percentage agreement (NPA) of 100% and 88.1%, respectively, compared to MC. All positive results (n/N = 9/17) were considered medically significant, with an increase in NPA to 100%. In purulent fluids/biopsies, BJIP and MC were concordant in 72% of cases (n/N = 18/25), with a per-sample PPA and NPA of 90% and 60%, respectively. For whole blood samples, almost all samples were negative by both methods (i.e., reference blood culture and BJIP), and the molecular test did not enable any further microbiological diagnosis. Conclusions: The BIOFIRE® Joint Infection Panel rapidly and accurately enabled or excluded a diagnosis of a POAI (~1 vs. 24–96 h for MC), optimising antimicrobial therapy.
The First Case of the Identification of a Microorganism Directly from Whole Blood Using MALDI-TOF Mass Spectrometry in an Onco-Hematological Pediatric Patient with Bloodstream Infection Venere Cortazzo, Maria del Carmen Pereyra Boza, Vanessa Tuccio Guarna Assanti, Gianluca Foglietta, Gianluca Vrenna, Marilena Agosta, Elena Chaiter, Martina Rossitto, Barbara Lucignano, Manuela Onori, Valeria Fox, Marco Becilli, Pietro Merli, Filippo Frioni, Carlo Federico Perno, Paola Bernaschi Antibiotics, 2025 Background: Bloodstream infections affect up to 20% of pediatric cancer patients receiving intensive care, contributing significantly to morbidity and mortality, with infection-related mortality rates reported to be as high as 16%. Methods: The identification of microorganisms directly from whole blood is difficult due to several factors, such as interference from host genomic material, low bacterial load, the endogenous components of whole blood and exogenous substances, which can interfere with the identification process. Nevertheless, rapid microbial diagnosis remains of paramount importance in these patients. Results and Conclusion: Here, we present the first case of bacterial pathogen identification directly from whole blood using MALDI-TOF mass spectrometry in an onco-hematological pediatric patient affected by sepsis and admitted to Bambino Gesù Children’s Hospital (IRCCS) in Rome, Italy.
A case of Ustilago spp. infection identified by whole genome sequencing in a pediatric patient undergoing open-chest extracorporeal membrane oxygenation Valeria Fox, Gianluca Vrenna, Gianluca Foglietta, Luna Colagrossi, Barbara Lucignano, Manuela Onori, Venere Cortazzo, Marilena Agosta, Martina Rossitto, Maria del Carmen Pereyra Boza, Vanessa Fini, Annarita Granaglia, Maria Giovanna Paglietti, Elisabetta Verrillo, Renato Cutrera, Paola Bernaschi, Carlo Federico Perno International Journal of Infectious Diseases, 2025 OBJECTIVES: Patients undergoing extracorporeal membrane oxygenation (ECMO) are susceptible to fungal infections, also from rare or emerging pathogens. We present a case of a 3-year-old male patient hospitalized for respiratory failure and subjected to open-chest ECMO support, with a fungal infection from a pathogen not identifiable by standard methods. METHODS: Although T2Candida panel (T2 Biosystems) resulted negative, blood cultures resulted positive for fungi after 4 days, confirmed by Gram staining. The fungus underwent typing using Bruker matrix-assisted laser desorption ionization-time of flight mass spectrometry system and Autobio Autof ms1000, which could not precisely identify the microorganism. Ultimately, whole genome sequencing (WGS) was performed directly on blood culture. RESULTS: WGS analysis revealed in 5 days the presence of a fungus belonging to the Ustilago genus, a group of fungi commonly found in the environment but rarely causing human diseases. CONCLUSIONS: To the best of our knowledge, we presented the first case of an Ustilago spp infection in a pediatric patient undergoing ECMO, not identified by standard techniques but only by WGS performed directly on a blood sample in 5 days. Despite the paucity of literature on Ustilago spp infections treatment, therapy adjustments led to the eradication of the pathogen, underscoring the importance of advanced molecular techniques for the correct and timely identification of these microorganisms.
Integrating Diagnostic Approaches in Infant Bacterial Meningitis Caused by a Non-K1 Escherichia coli: A Case Report Gianluca Vrenna, Marilena Agosta, Valeria Fox, Martina Rossitto, Venere Cortazzo, Serena Raimondi, Barbara Lucignano, Manuela Onori, Livia Mancinelli, Maria del Carmen Pereyra Boza, Vanessa Fini, Annarita Granaglia, Laura Lancella, Francesca Ippolita Calo’ Carducci, Costanza Tripiciano, Alberto Villani, Paola Bernaschi, Carlo Federico Perno Antibiotics, 2024
Acinetobacter baumannii Isolates from COVID-19 Patients in a Hospital Intensive Care Unit: Molecular Typing and Risk Factors Mariateresa Ceparano, Valentina Baccolini, Giuseppe Migliara, Claudia Isonne, Erika Renzi, Daniela Tufi, Corrado De Vito, Maria De Giusti, Maria Trancassini, Francesco Alessandri, Giancarlo Ceccarelli, Francesco Pugliese, Paolo Villari, Maria Angiulli, Stefania Battellito, Arianna Bellini, Andrea Bongiovanni, Lucilla Caivano, Marta Castellani, Monica Coletti, Alessia Cottarelli, Ludovica D’Agostino, Andrea De Giorgi, Chiara De Marchi, Irma Germani, Dara Giannini, Elisa Mazzeo, Shadi Orlandi, Matteo Piattoli, Eleonora Ricci, Leonardo Maria Siena, Alessandro Territo, Gianluca Vrenna, Stefano Zanni, Carolina Marzuillo Microorganisms, 2022
