Susan S. Chiu

Human coronavirus NL63 infection and other coronavirus infections in children hospitalized with acute respiratory disease in Hong Kong, China.

Abstract Background. Human coronavirus NL63 (HCoV-NL63) is a recently discovered human coronavirus found to cause respiratory illness in children and adults that is distinct from the severe acute respiratory syndrome (SARS) coronavirus and human coronaviruses 229E (HCoV-229E) and OC43 (HCoV-OC43). Methods. We investigated the role that HCoV-NL63, HCoV-OC43, and HCoV-229E played in children hospitalized with fever and acute respiratory symptoms in Hong Kong during the period from August 2001 through August 2002. Results. Coronavirus infections were detected in 26 (4.4%) of 587 children studied; 15 (2.6%) were positive for HCoV-NL63, 9 (1.5%) were positive for HCoV-OC43, and 2 (0.3%) were positive for HCoV-229E. In addition to causing upper respiratory disease, we found that HCoV-NL63 can present as croup, asthma exacerbation, febrile seizures, and high fever. The mean age (± standard deviation [SD]) of the infected children was 30.7 ± 19.8 months (range, 6–57 months). The mean maximum temperature (±SD) for the 12 children who were febrile was 39.3°C ± 0.9°C, and the mean total duration of fever (±SD) for all children was 2.6 ± 1.2 days (range, 1–5 days). HCoV-NL63 infections were noted in the spring and summer months of 2002, whereas HCoV-OC43 infection mainly occurred in the fall and winter months of 2001. HCoV-NL63 viruses appeared to cluster into 2 evolutionary lineages, and viruses from both lineages cocirculated in the same season. Conclusions. HCoV-NL63 is a significant pathogen that contributes to the hospitalization of children, and it was estimated to have caused 224 hospital admissions per 100,000 population aged ⩽6 years each year in Hong Kong.

Cytotoxic T Lymphocytes Established by Seasonal Human Influenza Cross-React against 2009 Pandemic H1N1 Influenza Virus

ABSTRACT While few children and young adults have cross-protective antibodies to the pandemic H1N1 2009 (pdmH1N1) virus, the illness remains mild. The biological reasons for these epidemiological observations are unclear. In this study, we demonstrate that the bulk memory cytotoxic T lymphocytes (CTLs) established by seasonal influenza viruses from healthy individuals who have not been exposed to pdmH1N1 can directly lyse pdmH1N1-infected target cells and produce gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α). Using influenza A virus matrix protein 1 (M158-66) epitope-specific CTLs isolated from healthy HLA-A2+ individuals, we further found that M158-66 epitope-specific CTLs efficiently killed both M158-66 peptide-pulsed and pdmH1N1-infected target cells ex vivo. These M158-66-specific CTLs showed an effector memory phenotype and expressed CXCR3 and CCR5 chemokine receptors. Of 94 influenza A virus CD8 T-cell epitopes obtained from the Immune Epitope Database (IEDB), 17 epitopes are conserved in pdmH1N1, and more than half of these conserved epitopes are derived from M1 protein. In addition, 65% (11/17) of these epitopes were 100% conserved in seasonal influenza vaccine H1N1 strains during the last 20 years. Importantly, seasonal influenza vaccination could expand the functional M158-66 epitope-specific CTLs in 20% (4/20) of HLA-A2+ individuals. Our results indicated that memory CTLs established by seasonal influenza A viruses or vaccines had cross-reactivity against pdmH1N1. These might explain, at least in part, the unexpected mild pdmH1N1 illness in the community and also might provide some valuable insights for the future design of broadly protective vaccines to prevent influenza, especially pandemic influenza.

Protective Efficacy of Seasonal Influenza Vaccination against Seasonal and Pandemic Influenza Virus Infection during 2009 in Hong Kong

BACKGROUND The relationship between seasonal influenza vaccine and susceptibility to 2009 pandemic A/H1N1 virus infection is not fully understood. METHODS One child 6-15 years of age from each of 119 households was randomized to receive 1 dose of inactivated trivalent seasonal influenza vaccine (TIV) or saline placebo in November 2008. Serum samples were collected from study subjects and their household contacts before and 1 month after vaccination (December 2008), after winter (April 2009) and summer influenza (September-October 2009) seasons. Seasonal and pandemic influenza were confirmed by serum hemagglutinination inhibition, viral neutralization titers, and reverse-transcription polymerase chain reaction performed on nasal and throat swab samples collected during illness episodes. RESULTS TIV recipients had lower rates of serologically confirmed seasonal A/H1N1 infection (TIV group, 8%; placebo group, 21%; P=.10) and A/H3N2 infection (7% vs 12%; P=A9), but higher rates of pandemic A/H1N1 infection (32% vs 17%; [Formula: see text]). In multivariable analysis, those infected with seasonal influenza A during the study had a lower risk of laboratory-confirmed pandemic A/H1N1 infection (adjusted odds ratio [OR], 0.35; 95% confidence interval [CI], 0.14-0.87), and receipt of seasonal TIV was unassociated with risk of pandemic A/H1N1 infection (adjusted OR, 1.11; 95% CI, 0.54-2.26). CONCLUSIONS TIV protected against strain-matched infection in children. Seasonal influenza infection appeared to confer cross-protection against pandemic influenza. Whether prior seasonal influenza vaccination affects the risk of infection with the pandemic strain requires additional study. CLINICAL TRIALS REGISTRATION ClinicalTrials.gov number NCT00792051 .

Comparison of nasopharyngeal flocked swabs and aspirates for rapid diagnosis of respiratory viruses in children

BACKGROUND The quality of clinical specimens is a crucial determinant for virological diagnosis. OBJECTIVES We compared the viral diagnostic yield for influenza A and respiratory syncytial virus (RSV) from the recently developed nasopharyngeal flocked swabs (NPFS) with nasopharyngeal aspirates (NPA) collected in parallel from 196 hospitalized children with acute respiratory infection during the peak period of influenza A and RSV activity in Hong Kong. Specimens were tested by RT-PCR for influenza A and RSV and viral load determined. They were also tested by direct immunofluorescence (DIF) for influenza A and B, RSV, parainfluenza types 1-3 and adenovirus. RESULTS Both NPA and NPFS had excellent sensitivity (100%) for detecting influenza A by RT-PCR but NPA was slightly more sensitive than NPFS for detecting RSV by both RT-PCR (100% vs. 92.3%) and DIF (87.2% vs. 84.6%) and for detecting influenza A by DIF (90.2% vs. 82.9%). Viral load for influenza A in NPA and NPFS was not significantly different but that for RSV was higher in NPA. CONCLUSION NPA remains the optimal specimen for diagnosis of respiratory infections by RT-PCR and DIF. However, collection of NPFS is easier to perform in an out-patient setting, was more acceptable to parents and less likely to generate aerosols than NPA engendering potentially less infection control hazard.

Rhinovirus infection in hospitalized children in Hong Kong: a prospective study.

Objectives: To analyze the clinical features and estimate the hospitalization disease burden of rhinovirus infection in children in Hong Kong. Methods: In this prospective study, nasopharyngeal aspirates were taken from children aged <18 years with symptoms of acute respiratory infection admitted to Queen Mary Hospital on one fixed day of the week during August 2001–July 2002 for detection of common respiratory viruses by immunofluorescence, viral culture, and for rhinovirus, human metapneumovirus, and coronaviruses by reverse transcription polymerase chain reaction. The clinical features of rhinovirus infections were analyzed and hospitalization disease burden was estimated. Results: Altogether 239 of the 426 nasopharyngeal aspirates (56.1%) were positive for respiratory viruses, including 151 patients with rhinovirus (35.4%). The median age was 2.34 years. Upper respiratory infection, asthma exacerbation, pneumonia, and acute bronchiolitis were diagnosed in 44.4%, 19.9%, 11.3%, and 7.9%, respectively. The most common symptoms were cough (81.5%), runny nose (76.8%), and fever (68.9%). Shortness of breath, wheezes, and crepitation were present in 25.8%, 29.1%, and 18.5%, respectively. Fifty-five of 99 patients (55.6%) had chest radiographic abnormalities, most commonly perihilar streakiness. Children with chronic diseases were more likely to have lower respiratory tract infection and these children required longer hospitalization (mean 0.6 days longer). Coinfection with other respiratory pathogens was common (33.1%). Conclusion: Rhinovirus is frequently associated with asthmatic exacerbations and lower respiratory tract infection, especially in children with chronic diseases and is potentially an important contributor to hospitalization in children in Hong Kong.

Early Diagnosis of Primary Human Herpesvirus 6 Infection in Childhood: Serology, Polymerase Chain Reaction, and Virus Load

Qualitative and quantitative polymerase chain reaction (PCR) for human herpesvirus 6 (HHV-6) DNA in whole blood and plasma was correlated with serology and clinical assessment in 143 children hospitalized for undifferentiated febrile illness to evaluate options for diagnosis of primary HHV-6 infection on the acute blood specimen. PCR and serology for HHV-7 were done in parallel to define serologic cross-reactions. Using HHV-6 seroconversion as the reference standard, detection of HHV-6 DNA in whole blood in the absence of antibody in the plasma was the most reliable evidence of primary HHV-6 infection. Detection of HHV-6 DNA in plasma and a high virus load in whole blood (>3.3 log10 copies/5 microL) had a sensitivity of 90% and 100%, respectively, in diagnosing primary HHV-6 infection. However, both were occasionally found in patients with other infections, possibly associated with HHV-6 reactivation. Maternal antibody may confound interpretation of serology in patients under 3 months of age.

Nasopharyngeal Carriage of Antimicrobial-Resistant Streptococcus pneumoniae among Young Children Attending 79 Kindergartens and Day Care Centers in Hong Kong

ABSTRACT Resistance to penicillin and multiple antimicrobial agents amongStreptococcus pneumoniae strains is becoming an increasing problem worldwide and in Asia. To determine the prevalence of carriage of S. pneumoniae isolates not susceptible to penicillin in young children, we obtained nasopharyngeal swab specimens from 1,978 children (ages, 2 to 6 years) attending 79 day care centers or kindergartens. Three hundred eighty-three strains of S. pneumoniae were isolated from these children. Fifty-eight percent of these isolates had reduced susceptibility to penicillin, 123 (32.1%) were intermediate, and 100 (26.1%) were resistant. A very high penicillin MIC (4 μg/ml) was found in 3.3% of the isolates. The isolates also demonstrated high rates of resistance to other antimicrobial agents (51.2% to cefaclor, 50.2% to cefuroxime, 42.8% to cefotaxime, 80.7% to trimethoprim-sulfamethoxazole, 77% to erythromycin, 60% to clindamycin, and 33.7% to chloramphenicol). No isolate was resistant to fluoroquinolone. Multidrug resistance (not susceptible to the β-lactams and three or more other classes) was found in 39.4% of the isolates. Risk factors for the carriage of S. pneumoniae not susceptible to penicillin were multiple physician visits in the preceding 3 months and use of antimicrobial agents by the individual or by household members in the preceding 3 months. In the logistic regression analysis, only the use of antimicrobial agents in the preceding 3 months was an independent risk factor (P = 0.004; odds ratio, 2; 95% confidence interval, 1.2 to 3.2). This study demonstrated the high prevalence of antibiotic-resistant S. pneumoniae in healthy young children in the community in Hong Kong.

Fecal carriage of CTXM type extended-spectrum beta-lactamase-producing organisms by children and their household contacts.

OBJECTIVES To investigate the epidemiology of fecal carriage of CTX-M type extended-spectrum beta-lactamases (ESBL)-producing organisms among children and their household contacts. METHODS Fecal carriage with CTX-M-producing organisms was studied in 53 children and 172 household members. Molecular methods were used to characterize the isolates. RESULTS The children were mostly healthy and hospitalized for relatively mild febrile illnesses. Overall, the prevalence of fecal carriage of CTX-M-producing bacteria was 43.5% (admission children, 37.7%; household children, 20.7% and household adults, 50.3%). Household colonization index (defined by number of household carriers/total number of members) was significantly higher among families with at least one individual having a history of prolonged (>3 months) out-of-town residence in the previous year (mean+/-standard deviation; yes group, 0.67+/-0.36 vs. no group, 0.39+/-0.28, P=0.009) and was inversely correlated with the living space per person (R-square=0.139, P=0.006). Among 29 households with at least two carriers of CTX-M-producing enterobacteria, six clusters of clonally related strains were shared by 15 individuals from seven households; with both intra- and inter-household transmission. CONCLUSION CTX-M beta-lactamases may spread extensively amongst family members in the home.

Fluoroquinolone and Other Antimicrobial Resistance in Invasive Pneumococci, Hong Kong, 1995–2001

Fluoroquinolone resistance among invasive pneumococci in Hong Kong was high and a result of clonal expansion and spread.

Increased Risk of Noninfluenza Respiratory Virus Infections Associated With Receipt of Inactivated Influenza Vaccine

Abstract We randomized 115 children to trivalent inactivated influenza vaccine (TIV) or placebo. Over the following 9 months, TIV recipients had an increased risk of virologically-confirmed non-influenza infections (relative risk: 4.40; 95% confidence interval: 1.31-14.8). Being protected against influenza, TIV recipients may lack temporary non-specific immunity that protected against other respiratory viruses.