A patient with respiratory toxigenic diphtheria in Greece after more than 30 years.

The introduction of treatment and systematic vaccination has significantly reduced diphtheria mortality; however, toxigenic strains continue to circulate worldwide. The emergence of an indigenous diphtheria case with fatal outcome in Greece, after 30 years, raised challenges for laboratory confirmation, clinical and public health management. Toxigenic Corynebacterium diphtheriae was isolated from an incompletely vaccinated 8-year-old boy with underlying conditions. The child passed away due to respiratory distress syndrome, before the administration of diphtheria antitoxin (DAT). All close contacts in family, school and hospital settings were investigated. Pharyngeal swabs were obtained to determine asymptomatic carriage. Chemoprophylaxis was given for 7 days to all close contacts and a booster dose to those incompletely vaccinated. Testing revealed a classmate, belonging to a subpopulation group (Roma), and incompletely vaccinated, as an asymptomatic carrier with an indistinguishable toxigenic strain (same novel multilocus sequence type, designated ST698). This case highlights the role of asymptomatic carriage, as the entry of toxigenic strains into susceptible populations can put individuals and their environment at risk. Maintenance of high-level epidemiological and microbiological surveillance, implementation of systematic vaccination in children and adults with primary and booster doses, availability of a DAT stockpile, and allowing timely administration are the cornerstone to prevent similar incidents in the future.

Distinction between rhinovirus-induced acute asthma and asthma-augmented influenza infection.

BACKGROUND: Rhinovirus (RV) is an established trigger of asthma attacks, whereas such a link is less consistent for influenza virus (IFV). OBJECTIVE: In the context of precision medicine, we hypothesized that IFV infection may cause a condition essentially different from RV, and we investigated this by evaluating clinical characteristics of RV/IFV-positive and -negative children with respiratory symptoms and/or fever. METHODS: One thousand two hundred and seven children, 6 months to 13 years old, hospitalized for flu-like illness were recruited in this cross-sectional study. Collected information included demographics, medical history, symptoms/physical findings/diagnosis at presentation and treatment. Nasal secretions were PCR-tested for IFV/RV. Associations were evaluated with adjusted logistic regression models. RESULTS: Rhinovirus positivity was associated with an asthma-like presentation, including increased wheeze/effort of breathing/diagnosis of acute asthma, and decreased fever/vomiting. Conversely, IFV+ children presented with less wheeze/effort of breathing/diagnosis of acute asthma, while they were more frequently febrile. In those with previous asthma history, both viruses induced wheeze; however, IFV was uniquely associated with a more generalised and severe presentation including fever, rales, intercostal muscle retractions and lymphadenopathy. These symptoms were not seen in RV+ asthmatics, who had fewer systemic signs and more cough. CONCLUSIONS AND CLINICAL RELEVANCE: In children with respiratory symptoms and/or fever, RV but not IFV is associated with wheeze and an asthma-like presentation. In those with an asthma history, IFV causes more generalised and severe disease that may be better described as "asthma-augmented influenza" rather than an "asthma attack." Differences in the acute conditions caused by these viruses should be considered in the design of epidemiological studies.

Community-associated Staphylococcus aureus pneumonia among Greek children: epidemiology, molecular characteristics, treatment, and outcome.

Staphylococcus aureus is an infrequent cause of community-associated (CA-SA) pneumonia in children. The aim of this study was to evaluate the clinical, epidemiological, microbiological, and molecular characteristics of CA-SA pneumonia among children hospitalized in two large tertiary care referral centers during an 8-year period. Cases of CA-SA pneumonia admitted between 2007 and 2014 were retrospectively examined through medical record review. Molecular investigation was performed for available strains; mecA, Panton-Valentine leukocidin (PVL) (lukS-lukF-PV), and fibronectin binding protein A (fnbA) genes were detected by polymerase chain reaction (PCR). Clones were assigned by agr groups, pulsed-field gel electrophoresis (PFGE), SCCmec, and multilocus sequencing typing (MLST). In total, 41 cases were recorded (boys, 61 %), with a median age of 4.3 months (range, 1-175). Methicillin-resistant S. aureus (MRSA) accounted for 31 cases (75.6 %). Complications included empyema (25/41, 61 %), pneumatoceles (7/41, 17 %), and lung abscess (1/41, 2.5 %). Intensive care unit (ICU) admission was required in 58.5 %. Two deaths occurred (4.9 %). Definitive therapy was based on vancomycin with or without other antibiotics (55.9 %), followed by clindamycin and linezolid (26.5 % each). All isolates were susceptible to vancomycin (MIC90 2 mg/L, range 1-2), teicoplanin, and linezolid, whereas 26.8 % were resistant to clindamycin. Among the 25 studied strains, 20 were mecA-positive (MRSA), carrying also the fnbA gene. Of these, 90 % belonged to the ST80-IV/agr3/PVL-positive clone. Methicillin-susceptible S. aureus (MSSA) strains showed polyclonality, 3/5 were PVL-positive, and 3/5 were fnbA-positive. MRSA and particularly the ST80-IV clone predominated among staphylococcal pneumonia cases in children. Treatment provided was effective in all but two patients, despite the relatively high minimum inhibitory concentration (MIC) of vancomycin and a high resistance to clindamycin.

Latent tuberculosis infection in children: diagnostic approaches.

Tuberculosis (TB) remains an important public health problem and a leading infectious cause of death. Diagnosis and treatment of latent tuberculosis infection (LTBI) is important for TB control and elimination. Nevertheless, the diagnosis of LTBI in both adults and children remains complex, since there is no gold standard. The development of interferon gamma release assays was a major breakthrough in the diagnosis of LTBI. The evaluation of IGRAs in the diagnosis of LTBI in children is proven to be difficult since childhood TB differs from adults as immune responses vary with age. Separate studies assessing IGRAs performance in children are still limited, and only a few of them divide results by narrow age groups Nevertheless, new approaches are being exploited by the ongoing research for the development of more efficient diagnostic tools. It is likely that many changes in both the diagnosis and management of LTBI will occur in the near future. We believe that better understanding of the immunopathology of latency can ultimately lead to the development of more effective strategies in TB control. In the present review we summarize current data on diagnosis of LTBI in children, underscoring the existing challenges and limitations.

Estimated strain coverage of serogroup B meningococcal vaccines: A retrospective study for disease and carrier strains in Greece (2010-2017).

Invasive meningococcal disease (IMD) is associated with high case fatality rates and long-term sequelae among survivors. Meningococci belonging to six serogroups (A, B, C, W, X, and Y) cause nearly all IMD worldwide, with serogroup B meningococci (MenB) the predominant cause in many European countries, including Greece (~80% of all IMD). In the absence of protein-conjugate polysaccharide MenB vaccines, two protein-based vaccines are available to prevent MenB IMD in Greece: 4CMenB (Bexsero™, GlaxoSmithKline), available since 2014; and MenB-FHbp, (Trumenba™, Pfizer), since 2018. This study investigated the potential coverage of MenB vaccines in Greece using 107 MenB specimens, collected from 2010 to 2017 (66 IMD isolates and 41 clinical samples identified solely by non-culture PCR), alongside 6 MenB isolates from a carriage study conducted during 2017-2018. All isolates were characterized by multilocus sequence typing (MLST), PorA, and FetA antigen typing. Whole Genome Sequencing (WGS) was performed on 66 isolates to define the sequences of vaccine components factor H-binding protein (fHbp), Neisserial Heparin Binding Antigen (NHBA), and Neisseria adhesin A (NadA). The expression of fHbp was investigated with flow cytometric meningococcal antigen surface expression (MEASURE) assay. The fHbp gene was present in-frame in all isolates tested by WGS and in 41 MenB clinical samples. All three variant families of fHbp peptides were present, with subfamily B peptides (variant 1) occurring in 69.2% and subfamily A in 30.8% of the samples respectively. Sixty three of 66 (95.5%) MenB isolates expressed sufficient fHbp to be susceptible to bactericidal killing by MenB-fHbp induced antibodies, highlighting its potential to protect against most IMD in Greece.

Performance of rapid influenza testing in hospitalized children.

Influenza infection is associated with high hospitalization rates among young children. Rapid diagnosis of influenza infection is particularly useful in order to prevent nosocomial infection and allows for the timely initiation of antiviral treatment. We evaluated the performance of a rapid influenza test in hospitalized children during the influenza season. All children (aged 6 months to 14 years) hospitalized with fever and/or respiratory symptoms, admitted during the 2005 influenza season, participated in the study. A multiplex reverse transcriptase polymerase chain reaction (RT-PCR), able to identify IFV-A H1N1, H3N2, and IFV-B subtypes, was performed on nasopharyngeal aspirates. The nasal swab was tested with a lateral-flow immunoassay (QuickVue Influenza Test). The performance of the rapid test was compared with the results of PCR. Influenza infection was diagnosed by PCR in 41/217 (19%) patients. Infection with influenza A virus (H3N2) was diagnosed in all cases. The performance of the QuickVue Influenza Test was estimated as follows: sensitivity 67.5%, specificity 96%, positive predictive value 79%, and negative predictive value 93%. The sensitivity of the test was higher in infants aged 6-12 months, in those with short duration of symptoms, and in the peak phase of the epidemic. The QuickVue Influenza Test is useful and reasonably accurate to detect influenza infection in hospitalized children during the influenza season. Infection with influenza virus is unlikely if the test is negative. A positive result suggests that infection is probable if influenza virus circulates in the community.

Scedosporium apiospermum skeletal infection in an immunocompetent child.

Abstract This is a case of Scedosporium apiospermum skeletal infection in a 10-year-old immunocompetent girl whose chief complaint was left knee swelling and pain. The child had a history of a bicycle accident two months before with a resultant deep penetrating trauma. Systematic administration of broad-spectrum antibiotics for 10 days was used, with no clinical improvement. Magnetic Resonance Imaging and arthrotomy of the affected joint revealed findings suggestive of osteomyelitis. Empirical intravenous antimicrobial therapy was instituted for a total of two months but one month after completion of antibacterial therapy the child returned to the hospital because of persistent knee swelling and pain. Following a new arthrotomy, Scedosporium apiospermum was isolated. The patient was cured with intravenous administration of voriconazole without any side effects and has no evidence of relapse after four years of follow-up.

Establishing nationally representative central line-associated bloodstream infection surveillance data for paediatric patients in Greece.

BACKGROUND: Healthcare-associated infections (HCAIs) are associated with increased morbidity and mortality and with excess costs. Central line-associated bloodstream infections (CLABSIs) are the most common HCAIs in neonates and children. AIM: To establish national benchmark data for rates of CLABSI in neonatal and paediatric intensive care units (NICUs and PICUs) and paediatric oncology units (ONCs). METHODS: Active surveillance for CLABSI was conducted from June 2016 to February 2017. A collaborative of 14 NICUs, four PICUs, and six ONCs participated in the programme. Surveillance definitions of central line (CL), central line utilization (CLU) ratio, CLABSI event, and CLABSI rate were based on the Centers for Disease Control and Prevention's 2014 National Healthcare Safety Network criteria. Medical records were assessed daily for calculating CL-days, patient-days, and susceptibility of isolated organisms. FINDINGS: A total of 111 CLABSI episodes were recorded. The overall mean CLABSI rate was 4.41 infections per 1000 CL-days, and the CLU ratio was 0.31. CLABSI rates were 6.02 in NICUs, 6.09 in PICUs, and 2.78 per 1000 CL-days in ONCs. A total of 123 pathogens were isolated. The most common pathogens were Enterobacteriaceae (36%), followed by Gram-positive cocci (29%), non-fermenting Gram-negative bacteria (16%), and fungi (16%). Overall, 37% of Gram-negative pathogens were resistant to third-generation cephalosporins and 37% to carbapenems. CONCLUSION: Nationally representative CLABSI rates were determined for paediatric patients. These data could be used to benchmark and serve as baseline data for the design and evaluation of infection control and antimicrobial stewardship interventions.

Meningococcal group C disease in Greece during 1993-2006: the impact of an unofficial single-dose vaccination scheme adopted by most paediatricians.

The aim of this study was to evaluate the impact of the meningococcal C conjugate vaccine on the epidemiology of meningococcal C disease in Greece. Data from the National Reference Laboratory for Meningococcal Disease and a questionnaire distributed to Greek paediatricians were assessed. Since the introduction of the vaccine in 2001, 72% of Greek paediatricians have administered it as one single dose to patients aged > or =12 months. This vaccination scheme has probably contributed to a dramatic decrease in the number of meningococcal C infections, which reached zero in 2004.

Management of child MDR-TB contacts across countries in the WHO European Region: a survey of current practice.

The World Health Organization European Region has one of the highest rates of multidrug-resistant tuberculosis (MDR-TB) in the world, resulting in many vulnerable children being exposed each year. Evidence for preventive therapy following MDR-TB exposure is limited and current guidance is conflicting. An internet-based survey was performed to determine clinical practice in this region. Seventy-two clinicians from 25 countries participated. Practices related to screening and decision-making were highly variable. Just over half provided preventive therapy for children exposed to MDR-TB; the only characteristic associated with provision was practice within the European Union (adjusted OR 4.07, 95%CI 1.33-12.5).

Variation in paediatric hospital antibiotic guidelines in Europe.

OBJECTIVE: To assess the availability and source of guidelines for common infections in European paediatric hospitals and determine their content and characteristics. DESIGN: Participating hospitals completed an online questionnaire on the availability and characteristics of antibiotic prescribing guidelines and on empirical antibiotic treatment including duration of therapy for 5 common infection syndromes: respiratory tract, urinary tract, skin and soft tissue, osteoarticular and sepsis in neonates and children. RESULTS: 84 hospitals from 19 European countries participated in the survey of which 74 confirmed the existence of guidelines. Complete guidelines (existing guidelines for all requested infection syndromes) were reported by 20% of hospitals and the majority (71%) used a range of different sources. Guidelines most commonly available were those for urinary tract infection (UTI) (74%), neonatal sepsis (71%) and sepsis in children (65%). Penicillin and amoxicillin were the antibiotics most commonly recommended for respiratory tract infections (RTIs) (up to 76%), cephalosporin for UTI (up to 50%) and for skin and soft tissue infection (SSTI) and bone infection (20% and 30%, respectively). Antistaphylococcal penicillins were recommended for SSTIs and bone infections in 43% and 36%, respectively. Recommendations for neonatal sepsis included 20 different antibiotic combinations. Duration of therapy guidelines was mostly available for RTI and UTI (82%). A third of hospitals with guidelines for sepsis provided recommendations for length of therapy. CONCLUSIONS: Comprehensive antibiotic guideline recommendations are generally lacking from European paediatric hospitals. We documented multiple antibiotics and combinations for most infections. Considerable improvement in the quality of guidelines and their evidence base is required, linking empirical therapy to resistance rates.

Interferon gamma inducible protein-10 in the diagnosis of paediatric tuberculosis infection in a low TB incidence country.

OBJECTIVE: To assess interferon-gamma inducible protein 10 (IP-10) as a diagnostic marker for tuberculous infection in children, particularly in relation to its differential diagnostic performance in young children. DESIGN AND RESULTS: A case-control study was conducted among 161 children and adolescents (mean age 6.3 years ± standard deviation 1.7; males n = 79, 49%). Fifty-four (33.5%) had active TB, 53 (33%) had latent tuberculous infection (LTBI), and 54 (33.5%) were non-LTBI controls. Unstimulated IP-10 levels did not differ between groups (P > 0.05 for all comparisons). TB-specific antigen stimulated IP-10 levels were more profoundly increased in infected groups than in controls (P < 0.001 for all comparisons). None of the IP-10-based diagnostic indexes demonstrated the ability to discriminate active disease from LTBI. A value of IP-10 ⩾ 1222 pg/ml had 83.3% sensitivity, 79.6% specificity, 80.4% positive predictive value and 82.7% negative predictive value for the diagnosis of LTBI. IP-10 based indexes demonstrated a trend towards better performance in the population group aged <5 years. CONCLUSION: The IP-10 assay could be useful in improving the diagnosis of LTBI in patients aged <5 years in combination with existing interferon-gamma release assays.

Pandemic influenza A vs seasonal influenza A in hospitalized children in Athens.

BACKGROUND: Data on pandemic H1N1 influenza (pH1N1) virus infection in hospitalised children are limited. AIMS AND OBJECTIVES: To examine the epidemiological and clinical characteristics of children hospitalised with pH1N1 at a large tertiary-care centre in Athens and compare them with those of children hospitalised with seasonal influenza A in previous years. METHODS: All children (n = 146) admitted with confirmed pH1N1 between October 2009 to February 2010 and January 2011 to May 2011 were included. Data on children ≧ 6 months of age (n = 109) were compared with those of 138 children admitted with seasonal influenza A who were examined during two previous influenza seasons (2002-2003 and 2004-2005). RESULTS: The age distribution was similar between seasonal and pandemic H1N1. Bronchial asthma was significantly more common in the seasonal influenza group but the clinical presentation was similar in the two groups, except that fever was more common in patients with pH1N1. Children admitted with seasonal influenza were more likely to develop acute otitis media. There were no significant differences between the two groups for severe outcomes (admission to the ICU, mechanical ventilation or death). Only one child with seasonal influenza (0.6%) and three with pH1N1 influenza (2%) required admission to the ICU. Mean length of hospitalisation was longer in the seasonal influenza group. CONCLUSION: Clinical manifestations were similar between pH1N1 and seasonal influenza, and the pandemic virus did not appear to cause more severe disease in hospitalised children.

Etiology of community-acquired pneumonia in hospitalized school-age children: evidence for high prevalence of viral infections.

BACKGROUND: Community-acquired pneumonia (CAP) in young children is most commonly associated with viral infections; however, the role of viruses in CAP of school-age children is still inconclusive. METHODS: Seventy-five school-age children hospitalized with CAP were prospectively evaluated for the presence of viral and bacterial pathogens. Nasopharyngeal washes were examined by polymerase chain reaction for viruses and atypical bacteria. Antibody assays to detect bacterial pathogens in acute-phase and convalescent-phase serum samples were also performed. RESULTS: A viral infection was identified in 65% of cases. Rhinovirus RNA was detected in 45% of patients; infection with another virus occurred in 31%. The most common bacterial pathogen was Mycoplasma pneumoniae, which was diagnosed in 35% of cases. Chlamydia pneumoniae DNA was not detected in any patient; results of serological tests were positive in only 2 patients (3%). Mixed infections were documented in 35% of patients, and the majority were a viral-bacterial combination. CONCLUSIONS: The high prevalence of viral and mixed viral-bacterial infections supports the notion that the presence of a virus, acting either as a direct or an indirect pathogen, may be the rule rather than the exception in the development of CAP in school-age children requiring hospitalization.

Prevalence and patterns of resistance of Streptococcus pneumoniae strains isolated from carriers attending day care centers in the area of Athens.

The prevalence and patterns of Streptococcus pneumoniae resistance to antibiotics was examined in 146 nasopharyngeal carrier strains obtained during April and May, 1997, from 382 healthy children attending eight day care centers (DCCs) in the area of Athens. Reduced susceptibility to at least one antibiotic was found in 32.6% as follows: penicillin 11.4% (intermediate), cefotaxime 0.8% (intermediate), trimethoprim-sulfamethoxazole 22.7%, erythromycin 13.6%, tetracycline 11.4%, chloramphenicol 8.3%. Most of the nonsusceptible to penicillin isolates belonged to serogroups 23, 9, and 19. Multidrug resistance was detected in 11.4% of S. pneumoniae isolates including five penicillin nonsusceptible serogroup 23 strains. More than half of the multidrug resistant strains were susceptible to penicillin and belonged to serogroups 6 (4), 23 (1), 19 (1), and 1(1). Strains that belonged to the same serogroup/serotype and had identical resistance patterns appeared to cluster in some DCCs. Antibiotic use in the previous month was associated with reduced susceptibility to penicillin (p = 0.007) and multidrug resistance (p = 0.012). In conclusion, a moderate prevalence of reduced susceptibility to penicillin in pneumococcal carrier strains was found in our community. Multidrug resistance was common and was often associated with susceptibility to penicillin. Several distinct patterns of resistance were observed, suggesting the spread of resistant clones to our country.

Group B streptococcus colonization of Greek pregnant women and neonates: prevalence, risk factors and serotypes.

OBJECTIVE: To evaluate the prevalence and risk factors of group B streptococcus (GBS) colonization among pregnant women and their neonates in Greece, and to examine the serotype distribution of the GBS strains isolated and their susceptibility to antibiotics. METHODS: A vaginal and a rectal swab were obtained from 1014 pregnant or parturient women followed at public and private hospitals in Athens and in a city of northern Greece. Cultures were also taken 24 h after birth from 428 neonates born to these women. RESULTS: The overall maternal and neonatal colonization rates were 6.6% and 2.4%, respectively. The vertical transmission rate was 22.5%. By logistic regression analysis, multiparity (>/=III) was associated with a lower colonization rate (odds ratio 4.4, 95% confidence interval 1.08-18.63). In contrast with other studies, middle-class women followed privately were more frequently colonized (10%) than those followed at the public hospital (3.9%) (odds ratio 3.1, 95% confidence interval 1.83-5.42). A higher number of prenatal visits was also associated with a higher colonization rate (change in true odds ratio when visits increased by one, 1.3; 95% confidence interval 1.14-1.60). No association was found between colonization and maternal age, previous obstetric history, marital status, nationality, prematurity, Caesarean section, or infant birth weight. The most common serotypes were II (26.9%), III (22.4%), Ia (19%), Ib (12%), and V (9%). A considerable proportion of the isolated strains was resistant to erythromycin (4.5%), clindamycin (6%), or both (6%). CONCLUSION: The rate and risk factors of maternal and neonatal GBS colonization may vary in different communities. These rates, as well as the incidence of neonatal disease, need to be thoroughly evaluated in each country to allow the most appropriate preventive strategy to be selected.

Otitis and respiratory distress episodes following a respiratory syncytial virus infection.

OBJECTIVE: To document, over two consecutive respiratory syncytial virus (RSV) seasons, the occurrence of acute otitis media (AOM) and recurrence of respiratory distress in children < 2 years of age hospitalized for respiratory distress. METHODS: Patients were examined during hospitalization and at 6 weeks and 6 months after discharge. RSV testing was performed on all patients, and hospitalized patients were evaluated daily for the occurrence of AOM. RESULTS: In total, 347 children were enrolled; 54.8% were RSV positive, and 45.2% were RSV negative. Children were most frequently diagnosed as having bronchiolitis (71.9%) or asthmatic bronchitis (17.9%); other diagnoses included pneumonia, laryngitis, and rhinitis. During hospitalization, AOM was diagnosed in 16.8% of RSV-positive versus 8.3% of RSV-negative children (P < 0.05). Six weeks after discharge, AOM was reported in 10.4% of RSV-positive as compared with 5.8% of RSV-negative patients. Six months later, AOM was reported in 2.9% of the RSV-positive and 7.6% of the RSV-negative patients. A second episode of acute respiratory distress, which either required (9) or did not require (35) hospitalization, occurred in 18.4% of the total population, with similar proportions of RSV-positive and RSV-negative children (17% versus 18.6%). CONCLUSION: We conclude that RSV appears to be an important contributing factor for the occurrence of AOM in young children hospitalized with respiratory distress. The occurrence of a second episode of acute respiratory distress did not appear to correlate with the previous RSV infection, but longer-term follow-up is required.

The impact of Greece's childhood tuberculosis screening programme on the epidemiological indexes in the greater Athens area.

SETTING: A hospital referral centre for childhood tuberculosis in Athens. OBJECTIVE: To evaluate the effectiveness of the screening programme implemented for childhood tuberculosis, through its impact on the epidemiological index. DESIGN: In Greece, tuberculosis has been systematically screened for in children since 1991 using the tuberculin skin test. The epidemiological and clinical profiles of all tuberculous children who attended the TB clinic were compared. The children were divided into those who attended in 1982-1990 and those who did so in 1991-1999. RESULTS: A total of 1122 TB patients were screened. In the second period there was an increase in numbers of immigrant children (3% vs. 28%, P = 0.0001), the rate of extra-pulmonary TB decreased (16% vs. 7.6%, P = 0.0001), patients identified by the screening programme increased (19% vs. 57%, P = 0.0001) and the number of symptomatic children fell (51% vs. 16%, P = 0.0001). The proportion of children who failed to attend for regular follow-up was lower during the second period (20% vs. 7%, P = 0.0001). CONCLUSIONS: Our study suggests that the screening programme applied in Greece during the last decade has contributed to the early identification of tuberculosis, and the limitation of symptomatic patients and extrapulmonary TB cases.