Diagnosing congenital cytomegalovirus infections using archived dried blood spots: A 15-year observational study, Portugal.

BACKGROUND: Cytomegalovirus (CMV) is a leading cause of congenital infections. Dried blood spots (DBS) collected in the first week of life (Guthrie cards) have been used in the diagnosis of CMV infection outside the three-week window period following birth. The present work summarizes the results of a 15-year observational study in which DBS from 1388 children were used for a late diagnosis of congenital CMV infection. METHODS: Three groups of children were studied: (i) symptomatic (with symptoms at birth or late sequelae) (N = 779); (ii) born to mothers with serological profile of primary CMV infection (N = 75); (iii) without any information (N = 534). A highly sensitive method of DNA extraction (heat-induced) from the DBS was used. CMV DNA was detected by a nested PCR. RESULTS: In total CMV DNA was detected in 7.5% (104/1388) of children. Symptomatic children showed a low rate of CMV DNA detection (6.7%) than children born to mothers with serological profile of primary CMV infection (13.3%) (p = 0.034). Sensorial hearing loss and encephalopathy were the two clinical manifestations with the highest CMV detection rate (18.3% and 11.1%, respectively). Children whose mothers had a confirmed primary infection showed a higher rate of CMV detection (35.3%) when compared with children whose mothers had a not confirmed primary infection (6.9%) (p = 0.007). CONCLUSION: The present work emphasises the importance of testing DBS in symptomatic children even a long time after symptoms onset and in children born to mothers with serologic diagnosis of maternal primary CMV infection when they miss the diagnosis during the three-week window following birth.

Genomic imbalances defining novel intellectual disability associated loci.

BACKGROUND: High resolution genome-wide copy number analysis, routinely used in clinical diagnosis for several years, retrieves new and extremely rare copy number variations (CNVs) that provide novel candidate genes contributing to disease etiology. The aim of this work was to identify novel genetic causes of neurodevelopmental disease, inferred from CNVs detected by array comparative hybridization (aCGH), in a cohort of 325 Portuguese patients with intellectual disability (ID). RESULTS: We have detected CNVs in 30.1% of the patients, of which 5.2% corresponded to novel likely pathogenic CNVs. For these 11 rare CNVs (which encompass novel ID candidate genes), we identified those most likely to be relevant, and established genotype-phenotype correlations based on detailed clinical assessment. In the case of duplications, we performed expression analysis to assess the impact of the rearrangement. Interestingly, these novel candidate genes belong to known ID-related pathways. Within the 8% of patients with CNVs in known pathogenic loci, the majority had a clinical presentation fitting the phenotype(s) described in the literature, with a few interesting exceptions that are discussed. CONCLUSIONS: Identification of such rare CNVs (some of which reported for the first time in ID patients/families) contributes to our understanding of the etiology of ID and for the ever-improving diagnosis of this group of patients.

The contribution of 7q33 copy number variations for intellectual disability.

Copy number variations (CNVs) at the 7q33 cytoband are very rarely described in the literature, and almost all of the cases comprise large deletions affecting more than just the q33 segment. We report seven patients (two families with two siblings and their affected mother and one unrelated patient) with neurodevelopmental delay associated with CNVs in 7q33 alone. All the patients presented mild to moderate intellectual disability (ID), dysmorphic features, and a behavioral phenotype characterized by aggressiveness and disinhibition. One family presents a small duplication in cis affecting CALD1 and AGBL3 genes, while the other four patients carry two larger deletions encompassing EXOC4, CALD1, AGBL3, and CNOT4. This work helps to refine the phenotype and narrow the minimal critical region involved in 7q33 CNVs. Comparison with similar cases and functional studies should help us clarify the relevance of the deleted genes for ID and behavioral alterations.

Prevalence and clinical features of adverse food reactions in Portuguese children.

BACKGROUND: The prevalence of adverse food reactions (AFR) has been increasing in the western world. Clinical manifestations are diversified and it may not be possible to clinically discriminate between IgE and non-IgE mediated AFR. In Portugal, the prevalence of AFR and food allergies in children is not known. Thus, the objectives of this study were to determine the prevalence of AFR in central Portugal. METHODS: Point prevalence study in 3-11 year-old schoolchildren from Central Portugal. Food-related questionnaires, skin prick tests (SPT) with foods and determination of food-specific IgE levels were performed. RESULTS: Of 4045 schoolchildren, 2474 (61.2%) accepted to be included in the study. Global prevalence of AFR was 7.1% (95% CI 6.2-8.1), based upon the initial questionnaire, 4.6% (95% CI 3.9-5.5), based upon a confirmatory questionnaire and the prevalence of probable food allergy (IgE-associated AFR: positive history + positive SPT and/or positive specific IgE) was 1.4% (95% CI 0.9-1.9). Most frequently implicated foods were fresh fruits, fish and egg. A first episode at an earlier age, mucocutaneous and anaphylactic reactions were more frequent in IgE-associated AFR. CONCLUSIONS: The prevalence of probable food allergy in 3-11 year old Portuguese children from central Portugal is low and parents over-report its frequency. Most frequently implicated foods were fresh fruit and fish. Immediate type, polysymptomatic, and more severe reactions may commence at an earlier age and be more frequent in IgE-associated than in non-IgE associated reactions.

Off-label drug prescribing in a Portuguese paediatric emergency unit.

BACKGROUND: Many drugs are prescribed outside the terms of the marketing authorization ("off-label"). Several studies have shown that this is a common practice in various European healthcare settings. OBJECTIVE: This study aimed to quantify and characterize off-label drug prescribing in children admitted to a Portuguese Paediatric Unit (PEU). SETTING: This study was conducted in the Paediatric Unit of the university teaching hospital of Cova da Beira Hospital Centre (CHCB), Covilhã, located in the Eastern Central Region of Portugal. METHOD: A descriptive study was conducted, including a sample of 700 children, randomly selected from those admitted between January to October 2010. Drug prescription was assessed by retrospective review of clinical files. MAIN OUTCOME MEASURE: Off-label prescribing was defined as the utilization of a drug at an indication, age, dosage, frequency or route of administration different from those recommended in the Summary of Product Characteristics (SPC). For purposes of this study only the medicines prescribed to be used after discharge from the hospital were studied. RESULTS: 364 boys and 336 girls, aged from 4 days to 18 years, were included in this study. Of the 724 medicines prescribed, 32.2 % were off-label. At least one drug was used off-label in 28.1 % of the studied population, corresponding to 46.1 % of the 427 children that received prescriptions. "Alteration in dosage" was the commonest reason for off-label prescribing (28.2 %). The off-label prescriptions comprised mainly drugs acting on the "Respiratory System" and "Anti-infectious agents for systemic use". Amoxicillin/clavulanic acid, paracetamol, amoxicillin, ibuprofen and salbutamol were the five active principles most frequently prescribed off-label. CONCLUSION: The prevalence of off-label drug prescribing in the Portuguese PEU of CHCB is high, the use in a dose or for an age group not approved in the SPC being the most common reasons for off-label prescription.

[Acute bronchiolitis: a prospective study]. / Bronquiolite aguda: estudo prospectivo.

INTRODUCTION: Bronchiolitis is the most common lower respiratory infection in children under 2 years old. Respiratory syncytial virus (RSV) is the most frequently involved etiologic agent. AIMS: To identify different viruses causing bronchiolitis and try to correlate them with demographic and clinical variables. To analyze diagnostic and therapeutic approache. METHODS: We conducted a prospective study, between November 2008 and March 2009 (5 months), including children < 2 years with bronchiolitis. Screening for RSV, parainfluenza 1-3 and adenovirus used immunofluorescence tests and screening for influenza A and B, human metapneumovirus (MPvh), human bocavirus (hBoV) and RSV used polymerase chain reaction (PCR) techniques. Data were analysed by using SPSS®. RESULTS: We included 78 children with 8.5 months mean age (83% < 12 months), 60% were male. The average duration of the disease was 15+5 days. Were on antibiotics 19.2%, because of concomitant acute otitis media (10.2%) or bacterial pneumonia (9%). 53% required hospital admission and the average length of stay was 7 days. Along hospitalization 95% of children required supplemental oxygen, 61% intravenous rehydration and 22% chest physiotherapy. Viral testing was positive in 59/75 children: RSV (69.3%), BoVh (22.7%), MPVh (4%), parainfluenza 3 (27%) and influenza A (2.6%). Co-infection with two viruses was detected in 23% of children. In 88% of children with positive samples for BoVh it has been detected RSV infection simultaneously. Children with co-infection (RSV + BoVh) required more often hospitalization compared with children infected with RSV alone (80% vs 60%, p=0.028), without significant differences in oxygen supplementation need and length of disease. CONCLUSIONS: RSV was the main etiologic agent and oxygen supplementation requirement justified the majority of hospitalizations. There was a high rate of co-infection with RSV and BoVh, but without longer disease. BoVh infection alone was uncommon.

Prevalence of latex sensitization and allergy in Portuguese children.

The prevalence of latex allergy has been increasing not only in risk groups but also in the general population, where it is accepted to average 1%. In children, latex sensitization prevalence studies are scarce and involve different population sampling and allergy testing methods, which makes it difficult to compare across studies. Nevertheless, existing studies point towards a low prevalence of latex allergy in children, which still needs to be confirmed in the Portuguese population. Aiming at studying the prevalence of latex sensitization and allergy in a sample of Portuguese children, we studied 182 children from two different hospital outpatient clinics. A standardized questionnaire focusing on atopic background, previous history and allergic signs or symptoms on exposure to latex or fruits was given to all children and parents. Skin prick testing was performed with a battery of common aeroallergens as well as latex. Serum total IgE, Phadiatop, F x 5E and latex-specific IgE were determined in all children. Specific IgE to latex-crossreacting fruits was determined in latex-sensitized children. Based upon the questionnaire, the prevalence of latex allergy would be 0.5%. The prevalence of latex sensitization would be 3.8%, when based solely upon skin prick testing, and 12.1% (>/=0.35 IU/ml) or 6.6% (>/=0.70 IU/ml) when based singly upon determination of latex-specific IgE. When positive results for either test were considered, the prevalence of latex sensitization was 14.3%. All latex-sensitized children were atopic. Sensitivity to latex-crossreacting foodstuffs was demonstrated in 61.5% of latex-sensitized children (16/26). This study shows that the prevalence of latex allergy and sensitization in Portuguese atopic and non-atopic children, as analysed using various diagnostic methods, is similar to that observed in other countries. In addition, the assessment of latex allergy and sensitization should always include skin prick testing and determination of serum IgE.