Marina Praprotnik

Human bocavirus and other respiratory viral infections in a 2-year cohort of hospitalized children.

Human bocavirus (HBoV) infection is reported worldwide and may cause severe respiratory tract infections. The aim of the present study was to assess the prevalence of HBoV, and other respiratory viral pathogens, in a 2‐year retrospective study of children admitted to hospital, and to investigate whether viral loads of HBoV DNA were associated with severity of infection. Between April 2007 and March 2009, 891 respiratory samples from 760 children admitted to hospital with acute respiratory tract infection were tested for the presence of respiratory viruses by real‐time PCR or direct immunofluorescence testing. HBoV DNA was detected by using internally controlled real‐time quantitative PCR assay and 25 samples selected at random were sequenced. The virus detected most frequently was rhinovirus, followed by respiratory syncytial virus, HBoV, and human metapneumovirus. HBoV DNA was detected in 18.4% of children admitted to hospital. HBoV was the only viral pathogen detected in 66/164 (40.2%) of HBoV DNA‐positive children and in 7.4% of all 891 samples. Ninety‐seven percent (64/66) of children with an HBoV single infection were diagnosed as having lower respiratory tract infection. Median HBoV DNA viral load was significantly higher in children when HBoV was detected as a single pathogen. Higher HBoV DNA viral loads were associated with prematurity and age. HBoV seems to be an important and frequent pathogen in respiratory tract infections in children, and it is likely that the severity of illness is comparable to the severity of RSV illness. J. Med. Virol. 84:99–108, 2011.

Prevalence, genotyping and macrolide resistance of Mycoplasma pneumoniae among isolates of patients with respiratory tract infections, Central Slovenia, 2006 to 2014

In this retrospective study we employed real-time polymerase chain reaction (PCR) to analyse the occurrence of Mycoplasma pneumoniae among upper and lower respiratory tract infections (RTI) in the Central Region of Slovenia between January 2006 and December 2014. We also used a culture and pyrosequencing approach to genotype strains and infer their potential macrolide resistance. Of a total 9,431 tested samples from in- and out-patient with RTI, 1,255 (13%) were found to be positive by M. pneumoniae PCR. The proportion of positive samples was 19% (947/5,092)among children (≤16 years-old) and 7% (308/4,339) among adults (>16 years-old). Overall, among those PCR tested, the highest proportions of M. pneumonia infections during the study period were observed in 2010 and 2014. In these two years, 18% (218/1,237) and 25% (721/2,844) of samples were positive respectively,indicating epidemic periods. From the 1,255 M. pneumoniae PCR-positive samples, 783 (614 from paediatric and 169 from adult patients) were successfully cultured. Of these, 40% (312/783) were constituted of strains belonging to the P1 type II genomic group, while 60% (469/783) contained strains of the P1 type I group. Two isolates comprised both P1 type Iand II strains. Results of a genotype analysis by year,showed that the dominant M. pneumoniae P1 type during the 2010 epidemic was P1 type II (82% of isolates;81/99), which was replaced by P1 type I in the 2014 epidemic (75%; 384/510). This observation could indicate that the two epidemics may have been driven by a type shift phenomenon, although both types remained present in the studied population during the assessed period of time. Only 1% of strains (7/783) were found to harbour an A2063G mutation in the 23S rRNA gene,which confers macrolide resistance, suggesting that the occurrence of M. pneumoniae macrolide resistance still seems to be sporadic in our geographic area.

First recovery of Rasamsonia argillacea species complex isolated in adolescent patient with cystic fibrosis in Slovenia – case report and review of literature

We report the isolation of the emerging fungal pathogen Rasamsonia aegroticola, which belongs Rasamsonia argillacea species complex, from a respiratory sample of a patient with cystic fibrosis. This filamentous fungus, resembling members of a Penicillium and Paecilomyces spp., was identified by morphology and confirmed by DNA sequence analysis. Susceptibility pattern showed high minimal inhibitory concentration of voriconazole and amphotericin B but low minimal inhibitory concentration of caspofungin, micafungin and itraconazole.

Autoimmune hepatitis as a presenting manifestation of mixed connective tissue disease in a child Case report and review of the literature

BackgroundLiver disease is rare in the course of mixed connective tissue disease. Most commonly liver steatosis or elevated liver function tests are reported and only a few cases of mixed connective tissue disease associated with autoimmune hepatitis were described.Case presentationWe report a case of an 11-year old boy with hepatitis on admission to the hospital and symptoms and signs of mixed connective tissue disease. Autoimmune hepatitis has been confirmed by liver biopsy.ConclusionTo the best of our knowledge this is the youngest patient with autoimmune hepatitis as a presenting manifestation of mixed connective tissue disease.

Clinical characteristics of infections caused by Mycoplasma pneumoniae P1 genotypes in children

Mycoplasma pneumoniae (M. pneumoniae) isolates can be classified into two major genetic groups, P1 type 1 (MP1) and P1 type 2 (MP2), based on the DNA sequence of the P1 adhesion protein gene. The aim of our study was to determine if M. pneumoniae P1 genotype is associated with disease manifestation and severity of acute M. pneumoniae infection. We compared epidemiological and clinical data of children infected with either MP1 or MP2. In addition, we separately analysed data of patients presenting with individual manifestations of M. pneumoniae infection. Data of 356 patients infected with MP1 were compared with those of 126 patients infected with MP2. MP2-infected children presented with higher median baseline C-reactive protein levels and were admitted to the hospital more often. The distribution of P1 genotype varied among groups of patients with different manifestations of M. pneumoniae infection. MP2 was more common than MP1 among patients with neurological and cardiovascular manifestations, whereas MP1 was more prevalent in other manifestations. The results from our large cohort indicate that the two P1 subtypes may have different pathogenic potential and that infections with MP2 strains could be more virulent than those with MP1 strains.

176 Elevation of 90 minutes plasma glucose during oral glucose tolerance testing predicts development of cystic fibrosis related diabetes and lung function decline

Objectives Recent studies have shown the effect of cystic fibrosis related diabetes (CFRD) on nutritional status and lung function of patients with CF. The aim of our study was to determine the prevalence of CFRD in Slovenian children with CF and the influence of hyperglycemia on their lung function and nutritional status. Methods Data from all Slovenian children with CF were collected. A diagnosis of CFRD was made based on oral glucose tolerance testing (OGTT) and continuous glucose monitoring. The highest level of blood glucose (BG) and the time during the OGTT were evaluated. Nutritional status and lung function of patients with CFRD were determined and compared to CF patients without CFRD. Results A total of 70 patients with a median age of 10.97 (0.9–22.3) years are managed in our Center. Twelve were diagnosed with CFRD (10 girls and 2 boys; median age 15.97 (9.21–18.59) years. The prevalence of CFRD was 17.1%. In 9 the BG levels >11.1 mmol/l at 90 ; during the OGTT were observed. The median value ob glycosylated hemoglobin (HbA1c) at diagnosis was 6.15 (5.6–6.8)%. The median FEV1 was 62% of predicted value and BMI 18.2 (12.4–21.7) kg/m 2 which is the 21 st percentile for age and gender. In patients with CFRD the median FEV1 was 24% lower compared with the rest of our CF patients while there was no significant difference in their BMI. Conclusions The prevalence of CFRD in our Center is comparable to the published data of CF patients of the same age group. Most of the CFRD patients had the highest BG level at 90 ; during OGTT. CFRD was associated with decline in lung function while it had no significant impact on nutritional status of our patients.

Cord blood interleukin-8 levels correlate with airway flow limitation at eight years of age in ex-very low birth weight infants

Background. Exposure to prenatal inflammation increases the risk for development of bronchopulmonary dysplasia. Aim. To evaluate the correlation between cord blood and gastric aspirate levels of interleukine-6 (IL-6) and interleukine-8 (IL-8) in preterm infants, and lung function at the age of 8 years. Methods. Between 2000-2002 we recruited 129 infants of gestational age < 30 wks. The concentration of IL-6 and IL-8 were measured in gastric aspirate and cord blood. At the age of 8 years, 30 ex-preterm infants, with mean gestational age of 27 wks and mean birth weight of 955 g, returned for pulmonary function measurement. To exclude major bias, a comparison between the study group and non-responder group was done and showed no statistically significant difference with respect to perinatal characteristics, ventilation days, bronchopulmonary dysplasia and cytokine concentration. Results. Pulmonary function test measurments in children born preterm were lower than in their term pairs. However, only the difference in forced mid-expiratory flow (FEF25-75%) was statisticaly significant. The concentration of IL-6 and IL-8 in cord blood and in gastric aspirate inversely correlated to all parameters of lung fuction at the age of 8 years, however only correlations between the concentration of IL-8 in cord blood and forced expired volume in one second/forced vital capacity (FEV1/FVC) (r = -0.38, p = 0.04) and FEF 25%-75% (r = -0.44, p = 0.02) were statistically significant. Conclusion. These results show a negative correlation between the concentration of IL-8 in cord blood and FEF25%-75% and FEV1/FVC, which suggests the important role of IL-8 in early airway remodeling.

151 Outbreak of multidrug-resistant Serratia marcescens detections in a paediatric cystic fibrosis (CF) centre

Introduction and Aims: Serratia marcescens (Sm) is an opportunistic pathogen in patients with cystic fibrosis (CF). CF patients acquire Sm infection mostly following manipulative airway procedures in hospitals, through patient-to-patient contacts or from the environment. This retrospective review aimed to present a case of an outbreak of Sm detections at a paediatric CF centre. Methods: Respiratory sampling, antibiotic susceptibility testing and antibiotic prescribing guidance were assessed. All isolates of Sm were additionally characterised by pulse field gel electrophoresis (PFGE) to determine the strain relatedness. Results: From April to July 2012 Sm was found in 5 patient’s airways’ samples (mean age 9.8 yrs, range 2−19 yrs, Centre size 63 patients, no Sm detected before). Three patients presented with pulmonary exacerbation and S. aureus, concomitantly. All received eradication therapy with TMP-SMX for 14 days based on antibiotic susceptibility. Treatment was repeated in 2 (40%) patients. Strict patient segregation and hygiene measures were implemented. The isolates of Sm did not differ in phenotypic and antibiotic susceptibility, but PFGE showed that all strains were biologically different, thus excluding cross-infection among patients or acquisition in the hospital. Patients therefore probably acquired Sm in a community-related setting. All patients were Sm free 10 months after first detection. Conclusions: Sm infection is of great concern due to its increasing resistance to antibiotics. Accurate identification is important in defining and managing outbreaks. PFGE can be a useful tool for understanding the molecular epidemiology.