Human bocavirus is a novel pathogen first detected in respiratory tract samples in 2005. People of different ages can be infected by human bocavirus. Children are the susceptible population, especially the infants aged from 6-24 months old. The epidemic season varies in different regions due to the differences in climate and geographical location, and it mainly occurs in autumn and winter. It's demonstrated that human bocavirus-1 is closely related to respiratory system diseases and even causes life-threatening critical illness. Also, the severity of symptom is positively correlated with viral load. Co-infections between human bocavirus-1 and other viruses often present high frequency occurrence. Human bocavirus-1 interferes immune function of host by inhibiting interferon secrete pathway. Currently, it remains limited knowledge and understanding of the roles of human bocavirus 2-4 in diseases, but the gastrointestinal diseases should be paid more attention. Detection of human bocavirus DNA by traditional polymerase chain reaction (PCR) assay shouldn't be regarded as conclusive diagnostic basis. Instead, combined with mRNA and specific antigen detection, it is beneficial to improve the accuracy of diagnosis. Till now, the knowledge of human bocavirus remains poorly studied, which is deserved to further progress.
Infant ; Humans ; Child ; Child, Preschool ; Human bocavirus ; Climate ; Coinfection ; Epidemics ; Interferons

Human bocavirus 1 (HBoV1) non-structural protein NS1 is a multifunctional protein important for virus replication and induction of apoptosis in host cell. To better understand the function of the NS1 protein, it is urgent to address reducing the toxicity of NS1 to host cells. In the present study, we established a stable cell line that regulates expression of NS1 of HBoV1. The recombinant lentivirus plasmid containing a regulatable promoter fused with ns1 gene was constructed and transfected into HEK 293T cells using transfection reagent. The HEK 293T cell lines stably expressing NS1-100 and NS1-70 proteins were established by screening resistant cells with puromycin and inducing NS1 expression with doxycycline. The expression of NS1 protein was determined by fluorescent labeling protein and Western blotting. HBoV1 promoter was transfected into stably expressing NS1 cell line and its trans-transcriptional activity was analyzed. The results showed that NS1 protein was expressed stably in the established cell lines and had a strong activation activity on the HBoV1 promoter driving luciferase gene. Taken together, this study provides a solid basis for further research on the function of NS1 and the pathogenesis of human bocavirus.
Human bocavirus ; Promoter Regions, Genetic ; Transcriptional Activation ; Viral Nonstructural Proteins ; Virus Replication

OBJECTIVE: Human bocavirus (HBoV) is a newly identified pathogen that can cause upper and lower respiratory infections usually in children; however, its clinical characteristics and significance in respiratory infections in adults have not been well known. Our objective was to evaluate the clinical features of respiratory HBoV infection and to describe the CT findings of HBoV pneumonia in adults. MATERIALS AND METHODS: A total of 185 adult patients diagnosed with HBoV infection at a tertiary referral center between January 2010 and December 2017 were retrospectively evaluated with respect to the clinical characteristics of HBoV infection and its risk factors for pneumonia. Chest CT findings for 34 patients with HBoV pneumonia without co-infection were analyzed and compared between immunocompetent (n = 18) and immunocompromised (n = 16) patients. RESULTS: HBoV infections were predominantly noted between February and June. Among the 185 patients with HBoV infection, 119 (64.3%) had community-acquired infections and 110 (59.5%) had pneumonia. In multivariable analysis, older age (odds ratio [OR], 1.02; 95% confidence interval [CI], 1.00–1.04; p = 0.045) and nosocomial infection (OR, 2.07; 95% CI, 1.05–4.10; p = 0.037) were associated with HBoV pneumonia. The main CT findings were bilateral consolidation (70.6%) and/or ground-glass opacities (64.7%); centrilobular nodules (14.7%) were found less frequently. The pattern of CT findings were not significantly different between immunocompetent and immunocompromised patients (all, p > 0.05). CONCLUSION: HBoV infection can be a potential respiratory tract infection in adults. The most frequent CT findings of HBoV pneumonia were bilateral consolidation and/or ground-glass opacities.
Adult ; Child ; Coinfection ; Community-Acquired Infections ; Cross Infection ; Human bocavirus ; Humans ; Immunocompromised Host ; Pneumonia ; Respiratory Tract Infections ; Retrospective Studies ; Risk Factors ; Tertiary Care Centers ; Tomography, X-Ray Computed

BACKGROUND: This study investigated the effect of fine dust concentrations in the air on the incidence of viral respiratory infections in the Republic of Korea.METHODS: A time series analysis using R statistics was performed to determine the relationship between weekly concentrations of fine dust in the air and the incidences of acute respiratory tract infections caused by the respiratory syncytial virus (RSV), adenovirus (HAdV), rhinovirus (HRV), human metapneumovirus (HMPV), human coronavirus (HCoV), human bocavirus (HBoV), human parainfluenza virus (HPIV), and influenza virus (IFV), from the beginning of 2016 to the end of 2017. Correlations between various meteorological factors and the amount of fine dust were analyzed using the Spearman's rank correlation coefficient. To analyze the relationship between viral infections and fine dust, a quasi-poisson analysis was performed.RESULTS: The incidence of the HAdV was proportional to fine dust and air temperature. The IFV was proportional to fine dust and relative humidity and was inversely proportional to temperature. The HMPV was proportional to fine dust, wind speed, and inversely proportional to relative humidity. The HCoV was proportional to micro dust, relative humidity, and inversely proportional to temperature. Both the HBoV and HPIV were directly proportional to fine dust, temperature, wind speed, and inversely proportional to relative humidity. The RSV was inversely proportional to fine dust, temperature, wind speed. A lag effect was observed for the influenza virus, in that its incidence increased 2–3 weeks later on the cumulative lag model.CONCLUSION: As the weekly average concentration of fine dust increases, the incidence of HAdV, HMPV, HCoV, HBoV, HPIV, and influenza increase.
Adenoviridae ; Air Pollution ; Coronavirus ; Dust ; Human bocavirus ; Humans ; Humidity ; Incidence ; Influenza, Human ; Metapneumovirus ; Meteorological Concepts ; Orthomyxoviridae ; Paramyxoviridae Infections ; Particulate Matter ; Republic of Korea ; Respiration Disorders ; Respiratory Syncytial Viruses ; Respiratory Tract Infections ; Rhinovirus ; Wind

OBJECTIVE: Newly identified human rhinovirus C (HRV-C) and human bocavirus (HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses. METHODS: A platform for culturing human airway epithelia in a three-dimensional (3D) pattern using Matrigel as scaffold was developed. The features of 3D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3D cells at designated time points were quantitated by real-time polymerase chain reaction (PCR). Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA. RESULTS: Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-1, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3D-cultured human airway epithelial (HAE) cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3D culture system. CONCLUSION Our data provide a preliminary indication that the 3D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV.
Collagen ; Drug Combinations ; Enterovirus ; growth & development ; isolation & purification ; Enterovirus Infections ; virology ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; virology ; Human bocavirus ; growth & development ; isolation & purification ; Humans ; Laminin ; Parvoviridae Infections ; virology ; Primary Cell Culture ; methods ; Proteoglycans ; Real-Time Polymerase Chain Reaction ; Respiratory Mucosa ; virology ; Virus Cultivation

PURPOSE: The aim of this study was to evaluate the severity of disease in children with acute bronchiolitis according to the type of infected virus. METHODS: From November 2007 to May 2015, 768 patients under 2 years of age who underwent real time-polymerase chain reaction of nasopharyngeal aspirates admitted to the Department of Pediatrics of Dongguk University Ilsan Hospital for acute bronchiolitis were enrolled. Severe bronchiolitis was defined as presence of one or more kinds among tachypnea, chest retraction, needs of O2 inhalation or ventilator care. RESULTS: The severity of bronchiolitis was increased with shorter fever duration (P < 0.001) and previous wheezing episodes (P = 0.005). In the case of single infection, respiratory syncytial virus (RSV) A only increased the severity of acute bronchiolitis (P = 0.012). However, the severity of illness decreased when RSV A coinfected with adenovirus (P = 0.034), human rhinovirus (P = 0.038), or human coronavirus NL63 (P = 0.042). On the other hand, when human rhinovirus was coinfected with enterovirus (P = 0.013) or parainfluenza 3 (P = 0.019), the severity was increased. When human metapneumovirus coinfected with human bocavirus, the severity was increased (P = 0.038). CONCLUSION: Acute bronchiolitis was associated with increased severity only when RSV A infected solely, but several viruses increased or decreased the severity when coinfection occurred. Therefore, it may be helpful in predicting the course of the acute bronchiolitis according to the affected virus.
Adenoviridae ; Bronchiolitis ; Child ; Coinfection ; Coronavirus NL63, Human ; Enterovirus ; Fever ; Hand ; Human bocavirus ; Humans ; Infant ; Inhalation ; Metapneumovirus ; Paramyxoviridae Infections ; Pediatrics ; Respiratory Sounds ; Respiratory Syncytial Viruses ; Rhinovirus ; Tachypnea ; Thorax ; Ventilators, Mechanical

Respiratory infections, which are caused by airborne pathogens, are the most common disease of all ages worldwide. This study was conducted to characterize the airborne respiratory pathogens in the public facilities in Busan, South Korea. A total of 260 public facilities were investigated in 2017, 52 seasonal indoor air from 2 hospitals and 208 indoor air samples from 208 randomly selected daycare centers. Among respiratory pathogen, 8 viral pathogens including human adenovirus (HAdV), human bocavirus (HBoV), human rhinovirus (HRV), human parainfluenza virus (HPIV), human respiratory syncytial virus (HRSV), human metapneumovirus (HMPV), human coronavirus (HCoV) and influenza virus (IFV), and 3 bacterial pathogens including Mycoplasma pneumoniae, Bordetella pertussis, and Chlamydophila pneumoniae, were investigated by multiplex real-time reverse transcription polymerase chain reaction. Pathogens were detected in 9 cases (3.4%). Among 9 positive samples, 6 (2.3%) cases were positive for HBoV and 3 (1.2%) cases were positive for IFV. All the positive cases were detected in daycare centers. Additionally, the concentration of HBoV was determined. In HBoV-positive samples, the cycle threshold (Ct) values of HBoV were 29.73~36.84, which are corresponding to the viral concentration of 4.91 × 10⁰ ~ 9.57 × 10² copies/ml. Serotype distribution of isolated HBoV was analyzed by sequencing of VP1/VP2 gene. All of the HBoV isolates were identified as HBoV type 1 with a high similarity among the isolates (>97%). No bacterial pathogen was identified in indoor air samples. Although virus concentration was not high in public facilities (daycare center), the presence of respiratory viral pathogens has been identified. Effective ventilation and air purification strategies are needed to reduce the indoor concentration of respiratory pathogens. A long-term and ongoing surveillance plan for respiratory pathogen management should be established.
Adenoviruses, Human ; Bordetella pertussis ; Busan ; Chlamydial Pneumonia ; Chlamydophila pneumoniae ; Coronavirus ; Human bocavirus ; Humans ; Korea ; Metapneumovirus ; Mycoplasma pneumoniae ; Orthomyxoviridae ; Paramyxoviridae Infections ; Pneumonia, Mycoplasma ; Polymerase Chain Reaction ; Public Facilities* ; Respiratory Syncytial Virus, Human ; Respiratory Tract Infections ; Reverse Transcription ; Rhinovirus ; Seasons ; Serogroup ; Ventilation

PURPOSE: It is well known that respiratory viral infection has epidemiological characteristics, including season. This study aimed to investigate the patterns of the prevalence of common respiratory viruses during a period of 10 years with regard to age, sex, and season in Korean children. METHODS: From June 2006 to November 2016, we obtained 11,798 specimens from patients aged less than 18 years who were admitted with lower respiratory infections. Ten respiratory viruses were detected using multiplex reverse transcription-polymerase chain reaction. RESULTS: Of 11,798 specimens, at least 1 virus was detected in 4,845 (41.1%). Respiratory syncytial virus (RSV, 18.9%) was the most common virus detected, followed by human rhinovirus (HRV, 14.8%), adenovirus (9.5%), and human bocavirus (HBoV, 7.4%). The detection rate of HRV was higher in male subjects (male 60.0% vs. female 40.0%, P=0.004), but the other viruses had no significant differences with regard to sex. The subjects who were positive for RSV test were youngest (median, 10.5 months; interquartile range, 3.0–25.0 months), followed by human coronavirus (median, 13.0 months), HRV (median, 14 months), and parainfluenza (median, 14 months). HBoV was most commonly detected in spring (29.3%), enterovirus in summer (25.8%), HRV in fall (22.6%), and RSV in October and winter (22.6%). CONCLUSION: We found that the prevalence of respiratory viruses in Korean children during a period of 10 years was associated with age, sex, and season when the infection occurred. Further nationwide data is warranted to infer clinical implication of our results.
Adenoviridae ; Child* ; Coronavirus ; Enterovirus ; Female ; Human bocavirus ; Humans ; Male ; Paramyxoviridae Infections ; Prevalence* ; Respiratory Syncytial Viruses ; Respiratory Tract Infections ; Rhinovirus ; Seasons*

PURPOSE: Croup, a common childhood respiratory illness with various severities, has many unanswered questions. Laryngotracheobronchopneumonitis (LTBP) is a disease entity considered to be an extension of croup to the lower respiratory tract. The object of this study was to compare epidemiology, clinical characteristics, and viral etiologic spectrum between croup and LTBP. METHODS: Patients hospitalized with croup at Gachon University Gil Hospital from January 2010 to April 2016 were recruited. LTBP was defined as pneumonia confirmed on radiographs of patients with croup. Clinical findings and demographic data were reviewed of patients whose nasopharyngeal swabs were done for viral analysis. RESULTS: A total of 371 patients with only croup and 63 patients with LTBP were included. Croup was found to be significantly associated with parainfluenza virus type 1 (P=0.006). LTBP was related to parainfluenza virus type 3, respiratory syncytial virus, and human bocavirus (P=0.001, P=0.030, and P=0.019, respectively). The duration of fever was longer in patients with LTBP than in those with croup (3.87±1.85 days vs. 2.86±1.80 days, P<0.001). CONCLUSION: Specific etiologic viruses might be associated with the progression from croup to LTBP. Pronged fever is also associated with progression from croup to LTBP.
Child* ; Croup* ; Epidemiology ; Fever ; Human bocavirus ; Humans ; Parainfluenza Virus 1, Human ; Parainfluenza Virus 3, Human ; Pneumonia ; Respiratory Syncytial Viruses ; Respiratory System

OBJECTIVETo investigate the prevalence of human bocavirus (HBoV) in children with acute lower respiratory tract infection and to explore the relationship between the viral load of HBoV and the clinical characteristics of acute lower respiratory tract infection in children.

METHODSA total of 1 554 nasopharyngeal aspirates from children who were hospitalized due to acute lower respiratory tract infection between March 2011 and March 2014 were collected. Quantitative real-time PCR was used to detect 12 RNA and 2 DNA viruses, adenovirus (ADV) and HBoV, and to measure the viral load of HBoV in HBoV-positive children. A comprehensive analysis was performed with reference to clinical symptoms and indicators.

RESULTSIn the 1 554 specimens, 1 212 (77.99%) were positive for viruses, and 275 (17.70%) were HBoV-positive. In HBoV-positive cases, 94.9% were aged <3 years, and there were more males than females. In the 275 HBoV-positive cases, 45 (16.36%) had single infection, and 230 (83.64%) had mixed infection. There was no significant difference in viral load between children with single infection and mixed infection (P>0.05). The patients with fever had a significantly higher viral load than those without fever (P<0.05). The children with wheezing had a significantly higher viral load than those without wheezing (P<0.05). There was no significant difference in viral load between children with mild, moderate, and severe acute lower respiratory tract infection (P>0.05).

CONCLUSIONSHBoV is one of the important pathogens of acute lower respiratory tract infection in children. Children with a higher viral load of HBoV are more likely to experience symptoms such as fever and wheezing. However, the severity of disease and mixed infection are not significantly related to viral load.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; Female ; Human bocavirus ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Tract Infections ; virology ; Viral Load