a Yu Chi

Challenges to Licensure of Enterovirus 71 Vaccines

Human enteroviruses usually cause self-limited infections except polioviruses and enterovirus 71 (EV71), which frequently involve neurological complications. EV71 vaccines are being evaluated in humans. However, several challenges to licensure of EV71 vaccines need to be addressed. Firstly, EV71 and coxsackievirus A (CA) are frequently found to co-circulate and cause hand-foot-mouth disease (HFMD). A polyvalent vaccine that can provide protection against EV71 and prevalent CA are desirable. Secondly, infants are the target population of HFMD vaccines and it would need multi-national efficacy trials to prove clinical protection and speed up the licensure and usage of HFMD vaccines in children. An international network for enterovirus surveillance and clinical trials is urgently needed. Thirdly, EV71 is found to evolve quickly in the past 15 years. Prospective cohort studies are warranted to clarify clinical and epidemiological significances of the antigenic and genetic variations between different EV71 genogroups, which is critical for vaccine design.

Long-term cognitive and motor deficits after enterovirus 71 brainstem encephalitis in children

OBJECTIVES. Several large outbreaks of enterovirus 71 infections have occurred in Taiwan during the past decade. Brainstem encephalitis was the most common neurologic complication. This study was designed to determine the long-term cognitive and motor outcomes of children with enterovirus 71 brainstem encephalitis. METHODS. We conducted a prospective follow-up study of children who met the case definition for enterovirus 71 brainstem encephalitis. Subjects were stratified into isolated brainstem encephalitis (stage II), autonomic nervous system dysregulation (stage IIIa), and pulmonary edema (stage IIIb). The subjects and their parents or guardians were interviewed using structured questionnaires and received comprehensive cognitive and neurologic examinations. Motor coordination, visual-motor skill, and intellectual ability were evaluated. RESULTS. Follow-up studies were conducted in 63 previously healthy children with enterovirus 71 brainstem encephalitis (49 stage II, 7 stage IIIa, and 7 stage IIIb). The mean time to follow-up was 2.8 ± 1.0 years (range: 1.4–4.9 years). Boys outnumbered girls by 3 to 2. The mean age at diagnosis was 2.4 ± 1.4 years (range: 0.3–7.1 years). The most common abnormal neurologic findings on admission were altered consciousness (47.6%), followed by abnormal activities of daily living (52.4%), cerebellar dysfunction (17.5%), and cranial nerve palsy (15.9%). At follow-up, 51 of 63 children had no detectable deficits. Among the remaining 12 children, 3 died during the follow-up. The remaining 9 children (14.3%) had residual deficits. Two of these with stage IIIb disease continued to have severe motor and respiratory failure. CONCLUSIONS. Residual defects were still present in a significant proportion of children with enterovirus 71 brainstem encephalitis at >2 years after their hospitalization. Children with stage II disease were most likely to have residual cerebellar defects. Those with stage IIIb disease continued to have severe respiratory and motor impairment. Long-term follow-up of this cohort is needed to determine the ultimate prognosis.

Clinical features of children infected with different strains of influenza B in southern Taiwan.

Background: This study was designed to determine the clinical characteristics of children infected with different strains of influenza B viruses isolated in southern Taiwan. The clinical features were compared with influenza A infection occurring in the same period. Methods: All children enrolled in the study had laboratory-confirmed infection with influenza A or B viruses. Influenza B speciation was performed by RNA extraction, cDNA synthesis, and amplification by polymerase chain reaction and sequencing. Demographic data, clinical findings, diagnoses, and outcomes were obtained. Results: During the study period, 163 strains of influenza A and 118 strains of influenza B were isolated. The Yamagata-like strains were most prevalent in 2001. New reassortant strains were identified since 2002 and became predominant in 2005 and 2006. Children with influenza B were more likely than those with influenza A to be diagnosed as upper respiratory tract infection, myositis, and gastroenteritis (P < 0.05). Children infected with Yamagata-like strains were more likely to develop lower respiratory tract infection (P < 0.05) and accounted for all cases of invasive disease. Children infected with the Victoria-like group had the longest hospital stays associated with severe bacterial superinfection. Conclusions: Currently new reassortant influenza B viruses are the predominant strains circulating in southern Taiwan. There is considerable similarity of clinical features between influenza A and B in children. The Yamagata-like strains were associated with more invasive infections. Continuous influenza virus surveillance is essential particularly in Taiwan where pandemic strains tend to appear earlier than in other countries.

Milrinone therapy for enterovirus 71-induced pulmonary edema and/or neurogenic shock in children: a randomized controlled trial.

Objective:Enterovirus 71-induced brainstem encephalitis with pulmonary edema and/or neurogenic shock (stage 3B) is associated with rapid mortality in children. In a small pilot study, we found that milrinone reduced early mortality compared with historical controls. This prospective, randomized control trial was designed to provide more definitive evidence of the ability of milrinone to reduce the 1-week mortality of stage 3B enterovirus 71 infections. Design:Prospective, unicenter, open-label, randomized, controlled study. Setting:Inpatient ward of a large tertiary teaching hospital in Ho Chi Minh City, Vietnam. Patients:Children (⩽18 yr old) admitted with proven enterovirus 71-induced pulmonary edema and/or neurogenic shock. Interventions:Patients were randomly assigned to receive intravenous milrinone (0.5 &mgr;g/kg/min) (n = 22) or conventional management (n = 19). Both groups received dopamine or dobutamine and intravenous immunoglobulin. Measurements and Main Results:The primary endpoint was 1-week mortality. The secondary endpoints included length of ventilator dependence and hospital stay and adverse events. The median age was 2 years with a predominance of boys in both groups. The 1-week mortality was significantly lower, 18.2% (4/22) in the milrinone compared with 57.9% (11/19) in the conventional management group (relative risk = 0.314 [95% CI, 0.12–0.83], p = 0.01). The median duration of ventilator-free days was longer in the milrinone treatment group (p = 0.01). There was no apparent neurologic sequela in the survivors in either group, and no drug-related adverse events were documented. Conclusions:Milrinone significantly reduced the 1-week mortality of enterovirus 71-induced pulmonary edema and/or neurogenic shock without adverse effects. Further studies are needed to determine whether milrinone might be useful to prevent progression of earlier stages of brainstem encephalitis.

High-Incidence of Human Adenoviral Co-Infections in Taiwan

Background Respiratory infections caused by adenovirus (HAdV) are common year round. Recently, a significant increase of adenoviral infections was observed in Taiwan. Objective To understand the prevalence and molecular epidemiology of respiratory adenovirus circulating in Taiwan for the past decade. Study Design One hundred and twenty-six human adenoviruses, isolated between 2002 to 2011, were characterized via DNA sequencing of the hexon and fiber genes. The nucleotide sequences were then compared by phylogenetic analysis. Results HAdV-B3 accounted for 64.3% (81/126) and peaked almost every year, whereas the sequences of hexon and fiber genes of HAdV-B3 were highly conserved in different years. A high incidence of co-infection of adenoviruses was observed (19.0%, 24/126); HAdV-B3 co-infected with HAdV-C2 was the most common combination (58.3%, 14/24). An additional interesting finding of repeated infection was noted in 10 children, all of whom showed first infection with adenovirus species HAdV-C, followed by species HAdV-B or HAdV-E. Conclusions HAdV-B3 was the predominant type of respiratory adenovirus circulating in Taiwan over the past ten years. This merits further attention for vaccine development. Furthermore, the observed high-incidence of adenoviral co-infections along with repeated infections found in our study provides important epidemiological insights into adenovirus infections.

Characterization of glycan binding specificities of influenza B viruses with correlation with hemagglutinin genotypes and clinical features

The carbohydrate binding specificities are different among avian and human influenza A viruses and may affect the tissue tropism and transmission of these viruses. The glycan binding biology for influenza B, however, has not been systematically characterized. Glycan binding specificities of influenza B viral isolates were analyzed and correlated to hemagglutinin (HA) genotypes and clinical manifestations. A newly developed solution glycan array was applied to characterize the receptor binding specificities of influenza B virus clinical isolates from 2001 to 2007 in Taiwan. Thirty oligosaccharides which include α‐2,3 and α‐2,6 linkage glycans were subjected to analysis. The glycan binding patterns of 53 influenza B isolates could be categorized into three groups and were well correlated to their HA genotypes. The Yamagata‐like strains predominantly bound to α‐2,6‐linkage glycan (24:29, 83%) while Victoria‐like strains preferentially bound to both α‐2,3‐ and α‐2,6‐linkage glycans (13:24, 54%). A third group of viruses bound to sulfated glycans and these all belonged to Victoria‐like strains. Based on the HA sequences, Asn‐163, Glu‐198, Ala‐202, and Lys‐203 were conserved among Victoria‐like strains which may influence their carbohydrate recognition. The viruses bound to dual type glycans were more likely to be associated with the development of bronchopneumonia and gastrointestinal illness than those bound only to α‐2,6 sialyl glycans (P < 0.05). Glycan binding analyses provide additional information to monitor the antigenic shift, tissue tropism, and transmission capability of influenza B viruses, and will contribute to virus surveillance and vaccine strain selection. J. Med. Virol. 84:679–685, 2012.

Prevalence and molecular characterization of staphylococcus aureus colonization among neonatal intensive care units in Taiwan

Background:Staphylococcus aureus, particularly methicillin-resistant (MRSA), is an important pathogen in neonatal intensive care units (NICUs). Carriage of S. aureus is a significant risk factor for subsequent infection. Objectives: To determine the current status of MRSA prevalence among NICU-hospitalized infants in Taiwan, we conducted this pilot island-wide survey. Methods: On two designated dates in 2011, each patient who stayed in the NICUs of 7 participating hospitals was included. Nasal and umbilical swabs were obtained and sent for detection of S. aureus. The prevalence and risk factors for MRSA carriage were analyzed. MRSA strains were tested for antimicrobial susceptibility and underwent molecular characterization. Results: A total of 251 subjects were included. The overall prevalence of S. aureus and MRSA carriage was 13 and 4.4%, respectively. Previous skin and soft tissue infection was the only predictor in multivariate analysis (OR 40.36; 95% CI 2.32-702.64; p = 0.011). Among 11 MRSA isolates, 3 pulsotypes were identified, with one major type (73%). Nine isolates carried a type IV staphylococcal chromosomal cassette, and 2 carried the type VT. All but one MRSA isolate belonged to linage sequence type 59, the community clone in Taiwan. Conclusions: On a designated date, 4.4% of the infants staying in NICUs in Taiwan carried almost genetically identical community strains of MRSA. MRSA colonization in these infants was significantly associated with previous skin and soft tissue infection.

Health Care-Associated Endocarditis Caused by Staphylococcus aureus with Reduced Susceptibility to Vancomycin

ABSTRACT Infective endocarditis (IE) caused by methicillin-resistant Staphylococcus aureus with reduced vancomycin susceptibility (SARV) has been reported worldwide. We report the successful treatment of a pediatric patient with SARV IE and characterization of the infecting strain. The MIC of vancomycin rose from 1.5 to 2 μg/ml, and the SARV was confirmed by population analysis.

Molecular Epidemiology and Clinical Manifestations of Adenovirus Respiratory Infections in Taiwanese Children.

AbstractHuman adenoviruses (HAdVs) are important causes of respiratory infections in children. They usually cause mild upper respiratory symptoms, but they can also produce severe pneumonia and other complications. The aims of this retrospective study were to better define the molecular epidemiology of respiratory adenoviruses circulating in Taiwanese children during 2002 and 2013, detect reinfections and co-infections, and characterize the clinical features and laboratory findings according to the causative genotypes.We collected a representative sample of 182 isolates of adenoviruses from 175 children during the 12-year study period. The most prevalent species was HAdV-B genotype 3 (HAdV-3) (92/182, 50.5%) followed by HAdV-C (HAdV-2) (38/182, 20.9%). A single outbreak of HAdV-E (6/182, 3.3%) was noted in 2007. The mean age of children with adenovirus infections was 3.7 ± 2.0 years, with a slight predominance of males (53.1%). Children with HAdV-B tended to be older, had more lower respiratory tract infections, gastrointestinal symptoms, and a higher rate of hospitalization than those with HAdV-C (P < 0.05). Adenovirus co-infections were noted in 25/175 (14.3%) of the children. The most frequent co-infections were with species B (HAdV-3) and C (HAdV-2) (14/25, 56.0%). Additional infections were noted in 23/175 (13.1%) of the children. Of these repeated infections, the initial isolates were always genotypes of HAdV-C. The second isolates were genotypes of HAdV-B or HAdV-E. The clinical features of the first HAdV-B infection and the reinfection of HAdV-B followed the HAdV-C were similar.In conclusion, HAdV-B, C, and E were the only adenovirus species that were isolated from children who were sufficiently ill with respiratory infections to require a visit to the hospital. Human adenovirus B (HAdV-3) accounted for half of these species. HAdV-B was more likely than other species to produce severe disease. The high incidence of adenovirus co-infection and reinfections with different HAdV species supports the need for continued surveillance and has major implications for development of vaccines.

Investigations of clinical isolations of oral poliovirus vaccine strains between 2000 and 2005 in southern Taiwan

BACKGROUND In Taiwan, trivalent oral poliovirus vaccine (tOPV) is in the routine immunization schedule, but its association with illnesses had not been examined. OBJECTIVES To investigate clinical presentations and viral characteristics of patients with poliovirus isolates. STUDY DESIGN Clinical data, vaccination records and viral sequences were retrospectively analyzed for patients from whom polioviruses were isolated during 2000-2005. RESULTS OPV-like strains were the only pathogen identified in 208 children who were diagnosed with lower respiratory tract infection (24.5%), acute gastroenteritis (16.8%) or upper respiratory tract infection (10.6%). Timing of poliovirus isolation relative to the tOPV vaccination was unusual in 59 patients, including 6 before any dose and 53 more than 10 weeks after the 3rd or later dose of tOPV. Sequence analyses of the VP1, 2C and 3C/D regions for 19 poliovirus isolates revealed that 4 had previously reported neurovirulence reversions, 1 had intertypic recombination, and 6 had both. No patient had neurological complications, but 3 died of myocarditis, including 2 with recombinant strains and 1 who never received OPV. CONCLUSION This study describes the isolation of OPV-like strains from patients with a variety of illnesses, raising concerns about their pathogenic potential in an area where tOPV is routinely administered. The detection of genetic variations among OPV-like strains warrants continuing surveillance for these variants in patients with severe illnesses besides neurological complications.