Giuseppe Indolfi

Long-Term Course of Chronic Hepatitis C in Children: From Viral Clearance to End-Stage Liver Disease

BACKGROUND & AIMS The natural course of chronic hepatitis C (CHC) in children is not well understood. The aim of this study was to assess the long-term course of CHC in a large sample of otherwise healthy children. METHODS From 1990 to 2005, 504 consecutive antihepatitis C virus (HCV)-positive children were enrolled at 12 centers of a national observatory and were followed up retrospectively/prospectively. RESULTS Putative exposure was perinatal in 283 (56.2%) cases, parenteral in 158 (31.3%), and unknown in 63 (12.5%). At baseline, 477 (94.6%) cases were HCV RNA seropositive, 118 (24.7%) of which were treated with standard interferon alpha. Ten years after putative exposure, the outcome in 359 HCV RNA-positive, untreated patients was (1) undetectable viremia in 27 (7.5%) (by Cox regression analysis, spontaneous viral clearance was independently predicted by genotype 3 [hazard ratio 6.44; 95% confidence interval: 2.7-15.5]) and (2) persistent viremia in 332 (92%) cases. Six of these 332 cases (1.8%) progressed to decompensated cirrhosis (mean age, 9.6 years). This latter group included 5 Italian children perinatally infected with genotype 1a (4 of the mothers were drug users). Thirty-three (27.9%) treated patients achieved a sustained virologic response. CONCLUSIONS Over the course of a decade, few children with chronic HCV infection cleared viremia spontaneously, and those who did were more likely to have genotype 3. Persistent viral replication led to end-stage liver disease in a small subgroup characterized by perinatal exposure, maternal drug use, and infection with HCV genotype 1a. Children with such features should be considered for early treatment.

Community-Acquired Bacteremic Pneumococcal Pneumonia in Children: Diagnosis and Serotyping by Real-Time Polymerase Chain Reaction Using Blood Samples

BACKGROUND The aim of this study was to use real-time polymerase chain reaction (RT-PCR) on blood samples to diagnose and serotype pneumococcal infection in a large cohort of Italian children hospitalized for community-acquired pneumonia. METHODS We conducted an observational study from April 2007 through June 2009 of children aged 0-16 years with a diagnosis of community-acquired pneumonia admitted to 83 pediatric hospitals in Italy. RESULTS Seven hundred fifty-three children were studied. RT-PCR found pneumococcal infection in 80 (10.6%) of 753 patients. In 292 patients, culture and RT-PCR were simultaneously performed. Streptococcus pneumoniae was identified in 47 of 292 patients; 45 (15.4%) tested positive by RT-PCR and 11 (3.8%) tested positive by culture. RT-PCR was significantly more sensitive than culture in revealing bacteremic pneumonia (odds ratio, 30.6; 95% confidence interval, 5.8-97.5; P<.001). Complicated pneumonia was found in 162 (21.5%) of 753 children; 152 (93.8%) of these 162 had parapneumonic effusion, and 51 (33.6%) had empyema. Children with complicated pneumonia were significantly older. Pneumococcal bacteremia was found by RT-PCR to occur significantly more frequently in children with complications (38 [23.5%] of 162) than in children with uncomplicated pneumonia (44 [7.4%] of 591; odds ratio, 3.8; 95% confidence interval, 2.30-6.30; P<.001). RT-PCR allowed serotyping from blood in 92.5% of patients. More than two-thirds of the pneumonia cases were due to nonpneumococcal conjugate vaccine 7 serotypes. Serotype 1 was the most frequent serotype (26 [32.5%] of 80) and was significantly associated with complications (50.0% in patients with complicated pneumonia vs 18.2% in patients with uncomplicated pneumonia; odds ratio, 4.5, 95% confidence interval, 1.48-14.03; P=.005) and older age. Serotype 19A was second in frequency (15.0%) and was significantly associated with younger age. CONCLUSIONS RT-PCR allows diagnosis and serotyping of pneumococcal bacteremic community-acquired pneumonia in children and is an important tool for evaluating serotype distribution in culture-negative samples.

Molecular detection methods and serotyping performed directly on clinical samples improve diagnostic sensitivity and reveal increased incidence of invasive disease by Streptococcus pneumoniae in Italian children

The aims of this study were to evaluate the incidence of invasive pneumococcal disease (IPD) in Italian children and perform serotyping by PCR-based assays directly on clinical samples. A 1-year paediatric (0–14 years) population-based surveillance study was designed to evaluate the incidence of IPD in the province of Florence, Italy, by cultural and molecular methods. Among 92 children (80 with pneumonia, 8 with meningitis/sepsis, 4 with arthritis), 4 cases of IPD were diagnosed both by culture and real-time PCR and 18 cases exclusively by molecular methods. The sensitivity of molecular methods was significantly higher than that of cultural methods (Cohen’s κ 0.41; McNemar P=0.000008). The incidence of IPD in children below 2 years of age was 11.5/100 000 and 51.8/100 000 by cultural and molecular methods, respectively. Pneumococcal serotyping by multiplex sequential PCR was obtained in 19/22 samples. Real-time PCR and multiplex sequential PCR can be used directly on biological samples, improving the ability to diagnose IPD. The incidence of IPD appears 5–10 times higher by PCR than by cultural methods.

Perinatal transmission of hepatitis C virus infection.

In industrialized countries, hepatitis C virus (HCV) is the most common cause of chronic liver disease in children. Perinatal transmission is the leading cause of infection. Perinatal transmission is confined almost always to women with detectable HCV ribonucleic acid (RNA) in the peripheral blood by the polymerase chain reaction but all children born to women with anti‐HCV antibodies should be tested for HCV. Some but not all studies found that a high concentration of serum HCV RNA is associated with a higher risk of transmission. Maternal peripheral blood mononuclear cell infection by HCV, membrane rupture of longer than 6 hr before delivery, and procedures exposing the infant to maternal blood infected with HCV during vaginal delivery are associated with an increased risk of transmission. Maternal coinfection with HCV and human immunodeficiency virus, maternal history of intravenous drug use and of HCV infection of the sexual partner of the mother predict the risk of perinatal transmission and are dependent on the peripheral blood mononuclear cell infection by HCV. Delivery by Cesarean section is not recommended in pregnant women infected with HCV. Infected mothers can breast feed safely their infants if the nipples are not damaged. A previous delivery of a child infected perinatally with HCV does not increase the risk of transmission in subsequent pregnancies. Immunogenetic factors and HCV genotypes are not related to HCV perinatal transmission. Despite an increased understanding of the risk factors involved in perinatal transmission of HCV, to date little is known about the transmission mechanisms and timing. J. Med. Virol. 81:836–843, 2009.

Realtime PCR Is More Sensitive than Multiplex PCR for Diagnosis and Serotyping in Children with Culture Negative Pneumococcal Invasive Disease

Background Pneumococcal serotyping is usually performed by Quellung reaction, considered the gold standard test. However the method cannot be used on culture-negative samples. Molecular methods can be a useful alternative. The aim of the study was to evaluate the use of Multiplex-sequential-PCR (MS-PCR) or Realtime-PCR on blood samples for diagnosis and serotyping of invasive pneumococcal disease (IPD) in a pediatric clinical setting. Methodology/Principal Findings Sensitivity and specificity of MS-PCR and Realtime-PCR have been evaluated both on 46 well characterized pneumococcal isolates and on 67 clinical samples from children with culture-negative IPD. No difference in sensitivity and specificity between MS-PCR and Realtime PCR was found when the methods were used on isolates: both methods could type 100% isolates and the results were always consistent with culture-based methods. On the contrary, when used on clinical samples 43/67 (64.2%) were typeable by MS-PCR and 61/67 (91.0%) by Realtime-PCR (p = 0.0004,K Cohen 0.3, McNemars p<0.001). Non-typeability by MS-PCR was associated in 18/20 cases (90.0%) with low bacterial load. The difference between the two methods was present both when they were used on normally sterile fluids (respectively 31/33 (93.9%) typeable samples for Realtime-PCR and 24/33 (72.7%) for MS-PCR, p = 0.047, 95%CL 0.03–0.98; K Cohen 0.3; McNemars p = 0.0016) and when they were used on nasopharyngeal swabs (respectively 30/34 (88.2%) typeable samples for Realtime-PCR and 19/34 (55.9%) for MS-PCR, p = 0.007, 95%CL 0.04–0.66); the presence of multiple pneumococcal serotypes in nasopharyngeal swabs was found more frequently by Realtime PCR (19/30; 63.3%) than by Multiplex-sequential PCR (3/19; 15.8%; p = 0.003;95%CL 1.87–39.97). Conclusions/Significance In conclusion, both MS-PCR and Realtime PCR can be used for pneumococcal serotyping of most serotypes/serogroups directly on clinical samples from culture-negative patients but Realtime-PCR appears more sensitive.

Comparison of the effect of antibiotic treatment on the possibility of diagnosing invasive pneumococcal disease by culture or molecular methods: A prospective, observational study of children and adolescents with proven pneumococcal infection

BACKGROUND Detection of Streptococcus pneumoniae in culture specimens in invasive pneumococcal disease (IPD) may be hampered by antibiotic treatment administered before hospital admission. Realtime polymerase chain reaction (RT-PCR) assays do not require viable bacteria and are therefore less influenced by antimicrobial therapy. It is not known how long results of culture or molecular tests remain positive after antibiotic therapy is begun. OBJECTIVE The goal of the current study was to assess, in a pediatric population with a diagnosis of IPD confirmed by laboratory tests (culture and/or RT-PCR assay), the relationship between use of antibiotic therapy before hospital admission and the result of diagnostic methods (culture or molecular techniques) after admission. METHODS This prospective, observational study was conducted from April 2006 through March 2009. All children and adolescents aged 0 to 16 years, admitted to the hospital with a diagnosis of IPD confirmed by culture and/or molecular methods, were included in the study. Previous antibiotic treatment (drug, duration of therapy) was recorded. Primers and probes designed from the pneumococcal autolysin gene (lytA) were used in an RT-PCR assay for detection of S pneumoniae. Antibiotic tolerability, permanent sequelae (after a 6-month follow-up), and deaths were recorded. RESULTS Eighty-three patients (50 males, 33 females; 80 white, 3 Asian; mean age, 4.6 years; median age, 4.0 years; age range, 10 days-16 years) were included in the study. Fifty-four patients presented with pneumonia, 26 with meningitis/sepsis (meningitis, 19; sepsis, 7), and 3 with arthritis. Results of RT-PCR assays were positive in all 83 patients (100.0%), and 28 of the 83 patients (33.7%) also had culture-positive findings. Forty-two of the 83 patients (50.6%) had received antibiotic treatment before hospital admission, and 41 (49.4%) had not received antibiotics. Results of cultures were positive in 9 of the 42 patients with IPD (21.4%) who had received antibiotic treatment and in 19 of the 41 patients with IPD (46.3%) who had not received antibiotics (odds ratio, 3.2; 95% CI, 1.1-9.3; P = 0.03). Molecular methods appeared more sensitive than culture in any type of disease studied but particularly in patients with pneumonia, in whom the difference was statistically significant (P = 0.043). The mean length of antibiotic therapy was 1.4 days (median, 1 day; SD, 0.53 day; range, 1-2 days) for culture-confirmed cases and 4.5 days (median, 4 days; SD, 3.08 days; range, 1-15 days) for cases confirmed by RT-PCR assay (P = 0.002). No adverse reactions to the antibiotics used during home or hospital treatment were found. Two patients with meningitis suffered permanent, severe neurologic sequelae, and 1 girl died of sepsis 3 days after hospital admission. No permanent sequelae were recorded in patients with pneumonia or arthritis. CONCLUSION In these children and adolescents with IPD, the molecular methods used appeared to be more sensitive than culture in any IPD patient, with a higher statistical significance in patients previously treated with antibiotics and in patients with pneumonia.

Thyroid function and anti-thyroid autoantibodies in untreated children with vertically acquired chronic hepatitis C virus infection

Objective  The reported data on thyroid function and anti‐thyroid autoantibodies in adults with untreated hepatitis C virus (HCV) infection are controversial. Data are scarce for HCV‐infected children, and only in those treated with interferon‐α (IFN‐α). We investigated thyroid function and anti‐thyroid autoantibodies in a cohort of untreated children with vertically acquired, chronic, HCV infection.

Cost of varicella-related hospitalisations in an Italian paediatric hospital: comparison with possible vaccination expenses

ABSTRACT Objective: The aims of the study were to evaluate the economic burden of hospitalisations due to varicella in an Italian paediatric hospital during a 1-year period and to compare the data with potential expenses projected for a varicella mass vaccination programme. Research design and methods: An observational, retrospective, cohort study was designed to measure hospital admission costs in a cohort of paediatric patients with varicella in a 12-month period. A cost comparison with a vaccination programme planned to prevent varicella in a whole birth cohort was performed. All children 0–16 years referred to the Anna Meyer Childrens Hospital (AMCH) were considered. Since AMCH is a tertiary-level hospital and accept patients from other Italian regions, in order to avoid overestimation of hospitalisation expenses, all analyses pertaining to both vaccination and hospitalisation costs were uniquely calculated on the basis of the cohort of residents in the district of Florence Results: A total of 279 children were examined in the emergency department for varicella; 47/279 (16.8%) were sent to the inpatient clinic. The highest rate of hospitalisation (85.1%) was found in children < 4 years of age, and the largest number of complications (87.2%) occurred in previously healthy children. Mean length of hospitalisation (5.7 ± 0.6 days) was similar to that reported in other western countries. Conclusion: Excluding any indirect cost, permanent sequelae and serious outcomes such as death, hospital expenses (corresponding to €239 654 in a 1-year period), would account for around 80% of total expenses for vaccinating an entire birth cohort (€310353).

Distribution of invasive meningococcal B disease in Italian pediatric population: Implications for vaccination timing

Neisseria meningitidis group B (MenB) is a leading cause of meningitis and sepsis. A new vaccine has been recently licensed. The aim of the present study was to evaluate the epidemiology of MenB disease in pediatric age and define the optimal age for vaccination. All patients aged 0-18 years admitted with a diagnosis of meningitis or sepsis to the 83 participating Italian pediatric hospitals were included in the study. Blood and/or cerebrospinal fluid (CSF) samples were tested by Realtime-PCR and/or culture. One hundred and thirty-six cases (mean age 5.0 years, median 2.7) of MenB disease were found. Among these, 96/136 (70.6%) were between 0 and 5 years, 61/136 (44.9%) were between 0 and 2 years. Among the latter, 39/61 (63.9%) occurred during the first year of life with highest incidence between 4 and 8 months. A case-fatality rate of 13.2% was found, with 27.8% cases below 12 months. Sepsis lethality was 24.4%. RT-PCR was significantly more sensitive than culture: 82 patients were tested at the same time by both methods, either in blood or in CSF; MenB was found by RT-PCR in blood or CSF in 81/82 cases (98.8%), culture identified 27/82 (32.9%) infections (Cohens Kappa 0.3; McNemars: p<10⁻⁵). The study shows that the highest incidence of disease occurs in the first year of age, with a peak between 4 and 8 months of life; 30% of deaths occur before 12 months. The results suggest that the greatest prevention could be obtained starting MenB vaccination in the first months of life; a catch-up strategy up to the fifth year of life could be considered. Our results also confirm that Realtime PCR is significantly more sensitive than culture. In those countries where only isolate positive infections are counted as cases, the incidence of MenB infection results highly underestimated.

Bilateral consolidation of the lungs in a preterm infant: an unusual central venous catheter complication

We describe a case of bilateral parenchymal consolidation with sudden respiratory distress in a preterm baby as a complication of peripherally inserted central catheter (PICC) dislocation. The X‐rays showed bilateral pulmonary consolidation with the catheter tip initially located in the right, and later in the left pulmonary artery. The catheter was withdrawn. As soon as the catheter was repositioned all clinical signs and symptoms disappeared. Neonatologists should consider the possibility of dramatic respiratory distress deriving from PICC dislocation. Careful tip catheter placement and conscientious monitoring may reduce morbidity.