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Mechanisms of interaction between Bordetella pertussis and other viral agents are yet to be fully explored. We studied the inflammatory cytokine expression patterns among children with both viral-bacterial infections. Nasopharyngeal aspirate (NPA) samples were taken from children, aged < 1 year, positive for Rhinovirus, Bordetella pertussis and for Rhinovirus and Bordetella pertussis. Forty cytokines were evaluated in NPA by using human cytokine protein arrays and a quantitative analysis was performed on significantly altered cytokines. Forty cytokines were evaluated in NPA by using human cytokine protein arrays and a quantitative analysis was performed on significantly altered cytokines. Our results show that co-infections display a different inflammatory pattern compared to single infections, suggesting that a chronic inflammation caused by one of the two pathogens could be the trigger for exacerbation in co-infections.

Background: Respiratory syncytial virus (RSV) is a leading cause of acute respiratory infections (ARIs) in children. However, the clinical impact of RSV co-infection with other respiratory viruses remains unclear. This study investigates the frequency and clinical outcomes of RSV infections in a large paediatric cohort. Methods: Paediatric patients with RSV-positive respiratory samples admitted to Bambino Gesù Children’s Hospital between January 2022 and April 2024 were analysed. Results: Within 17,259 respiratory samples from 9877 paediatric patients, 952 (9.6%) were RSV-positive. Among these, 637 patients with ARI were included. RSV affected the lower respiratory tract in 549 cases (86.2%) and the upper tract in 88 (13.8%) cases. RSV mono-infection was found in 286 (44.9%) patients, while 351 (55.1%) patients had co-infections. Mono-infections showed lower cycle-threshold (CT) than co-infections in both upper and lower tract (p-value:0.002 and 0.037, respectively). Pneumonia was associated with RSV co-infection (N = 48, 15.4%), whereas bronchiolitis was mostly seen in mono-infection (N = 196, 78.1%, p-value:0.002). Regression analysis showed an association between pneumonia and co-infection (AOR: 1.97 [1.06–3.64], p-value = 0.031), higher CT (AOR [95% CI]: 1.07 [1.02–1.11], p-value = 0.006) and older age (AOR [95% CI]: 1.48 [1.31–1.68], p-value < 0.001), whereas bronchiolitis was associated with mono-infection, younger age and lower CT. Conclusions: This study highlights the role of RSV in paediatric disease and emphasises the importance of early diagnosis, personalised treatment and preventive strategies to improve outcomes and reduce the burden of disease.

Human adenoviruses (HAdV) comprise more than 100 genotypes with species-specific differences in tropism and immune response and can cause severe infections in immunocompromised patients. This study aimed to characterise the HAdV species involved in pediatric infections to assess their clinical impact and guide future therapeutic strategies based on AdV-specific T-cell responses. Between January and October 2024, 595 pediatric HAdV diagnoses were made at the Bambino Gesù Children’s Hospital (Rome), and whole-genome sequencing was performed on 60 samples. Most patients (91.7%) were hospitalised, including both immunocompetent (75%) and immunocompromised (25%) children. Gastrointestinal and respiratory symptoms were more common in immunocompetent patients, whereas immunocompromised patients experienced longer hospitalisations and persistent viral infections. Species F (F41) was most prevalent (63.3%), especially among immunocompetent patients, while species C and A predominated in immunocompromised children, with species A associated with severe disease. Viral loads were significantly higher for species F than for species A and C, independent of immune status. Co-infections were frequent (63.3%), with species C particularly linked to them. In conclusion, HAdV distribution differed by immune status, with species F predominating in immunocompetent children and species C and A more common in immunocompromised patients. Whole-genome sequencing may enhance surveillance, enable earlier diagnosis, and support the development of genotype-specific immunotherapies.

Viral co-infections are frequently observed among children, but whether specific viral interactions enhance or diminish the severity of respiratory disease is still controversial. This study aimed to investigate the type of viral mono- and co-infections by also evaluating viral correlations in 3525 respiratory samples from 3525 pediatric in/outpatients screened by the Allplex Respiratory Panel Assays and with a Severe Acute Respiratory Syndrome-COronaVirus 2 (SARS-CoV-2) test available. Overall, viral co-infections were detected in 37.8% of patients and were more frequently observed in specimens from children with lower respiratory tract infections compared to those with upper respiratory tract infections (47.1% vs. 36.0%, p = 0.003). SARS-CoV-2 and influenza A were more commonly detected in mono-infections, whereas human bocavirus showed the highest co-infection rate (87.8% in co-infection). After analyzing viral pairings using Spearman’s correlation test, it was noted that SARS-CoV-2 was negatively associated with all other respiratory viruses, whereas a markedly significant positive correlation (p < 0.001) was observed for five viral pairings (involving adenovirus/human bocavirus/human enterovirus/metapneumoviruses/rhinovirus). The correlation between co-infection and clinical outcome may be linked to the type of virus(es) involved in the co-infection rather than simple co-presence. Further studies dedicated to this important point are needed, since it has obvious implications from a diagnostic and clinical point of view.

Since late 2023, an increase in Bordetella pertussis infections has been noticed in Europe, particularly among children. Our data showed the upward trend of B. pertussis cases in the Lazio region, even among adults with severe influenza-like illnesses, highlighting the necessity for maintaining high vaccination rates across both children and adults. These findings underscore the urgent need for clinicians to maintain a high index of suspicion for B. pertussis in patients with respiratory symptoms, prioritize nasopharyngeal swabs for accurate diagnosis, assess for co-infections, verify booster vaccination status in adults, and support timely reporting to public health authorities.

Background In recent months, Bordetella pertussis has reappeared after maintaining a low rate for many years. Although pertussis is usually characterized by a favorable course, several factors can contribute to the severity of the disease, such as mixed respiratory infections. In this study, we evaluate B.pertussis cases observed in the pediatric population followed at the Bambino Gesù Children's Hospital and analyzed the potential impact of co-infections in relation to disease severity. Methods From January to May 2024, a total of 1,151 children and adolescents (both inpatients and outpatients) were screened for the presence of respiratory pathogens, including B.pertussis, with clinically relevant respiratory symptoms. Results Among the 1,151 patients screened, 66 tested positive for B.pertussis . Fourteen patients had respiratory failure, and six of them required intensive care unit (ICU) admission, while 52 had mild infection. 23.3% of patients had B.pertussis alone, while 76.7% had co-infections (including 5 patients admitted to the ICU). A higher co-infection rate was observed in patients with respiratory failure than in those without failure (92.9% vs. 69.0%, p -value:0.041). Rhinovirus, Metapneumovirus and Parainfluenza-virus were the most prevalent in our pediatric population. Co-infections of human bocavirus with B.pertussis were observed exclusively in patients with respiratory failure. Conclusions Our results highlighted an increase in B.pertussis cases from January to May 2024, reaching a peak of cases in the month of May. This study shows a high rate of B.pertussis co-infection, and a trend toward association between B.pertussis and specific viruses, that might play a role in increasing disease severity.

Background Clinical criteria for pertussis diagnosis and clinical case definitions for surveillance are based on a cough lasting two or more weeks. As several pertussis cases seek care earlier, a clinical tool independent of cough duration may support earlier recognition. We developed a data-driven algorithm aimed at predicting a laboratory confirmed pertussis. Methods We enrolled children <12 months of age presenting with apnoea, paroxistic cough, whooping, or post-tussive vomiting, irrespective of the duration of cough. Patients underwent a RT-PCR test for pertussis and other viruses. Through a logistic regression model, we identified symptoms associated with laboratory confirmed pertussis. We then developed a predictive decision tree through Quinlan's C4.5 algorithm to predict laboratory confirmed pertussis. Results We enrolled 543 children, of which 160 had a positive RT-PCR for pertussis. A suspicion of pertussis by a physician (aOR 5.44) or a blood count showing leukocytosis and lymphocytosis (aOR 4.48) were highly predictive of lab confirmed pertussis. An algorithm including a suspicion of pertussis by a physician, whooping, cyanosis and absence of fever was accurate (79.9%) and specific (94.0%) and had high positive and negative predictive values (PPV 76.3% NPV 80.7%). Conclusions An algorithm based on clinical symptoms, not including the duration of cough, is accurate and has high predictive values for lab confirmed pertussis. Such a tool may be useful in low resource settings where lab confirmation is unavailable, to guide differential diagnosis and clinical decisions. Algorithms may also be useful to improve surveillance for pertussis and anticipating classification of cases.

Background Respiratory syncytial virus (RSV) infection in children under 5 years have a significant clinical burden, also in primary care settings. This study investigates the epidemiology and burden of RSV in Italian children during the 2019/20 pre-pandemic winter season. Methods A prospective cohort study was conducted in two Italian regions. Children with Acute Respiratory Infection (ARI) visiting pediatricians were eligible. Nasopharyngeal swabs were collected and analyzed via multiplex PCR for RSV detection. A follow-up questionnaire after 14 days assessed disease burden, encompassing healthcare utilization and illness duration. Statistical analyses, including regression models, explored associations between variables such as RSV subtype and regional variations. Results Of 293 children with ARI, 41% (119) tested positive for RSV. Median illness duration for RSV-positive cases was 7 days; 6% required hospitalization (median stay: 7 days). Medication was prescribed to 95% (110/116) of RSV cases, with 31% (34/116) receiving antibiotics. RSV subtype B and regional factors predicted increased healthcare utilization. Children with shortness of breath experienced a 36% longer illness duration. Conclusions This study highlights a significant clinical burden and healthcare utilization associated with RSV in pre-pandemic Italian primary care settings. Identified predictors, including RSV subtype and symptomatology, indicate the need for targeted interventions and resource allocation strategies. RSV epidemiology can guide public health strategies for the implementation of preventive measures.

Ageing is associated with a progressive decline and remodelling of the immune system. Also, the efficacy of COVID-19 vaccines has been observed to depend on subjects’ age. The post-vaccination data about patients aged > 90 years old is scarcely represented in the literature. The antibody titre profiles of elderly vaccinated subjects (age > 90 years old) were evaluated and compared with profiles obtained in a younger population (age 23–69 years old). To the best of our knowledge, this is the first report providing post-vaccination serological data in subjects aged 90 + years old. This study suggests that distinct SARS-CoV-2 viral-specific antibody response profiles vary based on anti-N serostatus, age, and sex in the very elderly adults. The data obtained could impact the organisation of the vaccination campaign (i.e., prioritisation strategies, administration of additional doses) and the factors that facilitate intentions to receive the vaccination among elderly adults (i.e., vaccine effectiveness).

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.