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      Human Bocavirus in Infants, New Zealand


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          To the Editor: In 2005, a parvovirus, subsequently named human bocavirus (HBoV), was discovered in respiratory samples taken from infants and children hospitalized at Karolinksa University Hospital, Sweden, with lower respiratory tract infection ( 1 ). HBoV has since been identified in infants and children with respiratory illness in >17 countries, at frequencies ranging from 1.5% to >18.0%. In the past decade New Zealand has experienced increasing bronchiolitis hospitalization rates, currently >70 admissions per 1,000 infants. To determine the contribution of HBoV to New Zealand’s bronchiolitis disease prevalence, we tested samples collected from infants hospitalized with community-acquired bronchiolitis ( 2 ) during 3 consecutive winter epidemics (June to October, 2003; July to October, 2004; and June to October, 2005) in Wellington, NZ, for HBoV by PCR. The Central Regional Ethics Committee approved the study. Written, informed consent was obtained from the parent or guardian. Demographic, clinical, and laboratory data were collected during hospitalization. Ethnicity of those who ascribe to >1 group was determined by using a national census method that prioritizes ethnicity as follows: Māori>Pacific>Other>New Zealand European. Oxygen requirement was determined to be the best measure of bronchiolitis severity ( 2 ). Infants who needed assisted ventilation or continuous positive airway pressure were classified severe; those who required oxygen supplementation, moderate; and infants who were hospitalized but did not require supplemental oxygen, mild. Nucleic acid was extracted from thawed nasopharyngeal aspirates (stored at 80°C) by using a High Pure Viral Nucleic Acid kit (Roche Diagnostics, Auckland, NZ). The HBoV nonstructural protein (NP-1) gene was amplified by using primers 188F (5′-GAGCTCTGTAAGTACTATTAC-3′) and 542R (5′-CTCTGTGTTGACTGAATACAG-3′) ( 1 ) with Expand High Fidelity DNA Polymerase (Roche Diagnostics, Basel, Switzerland) for 35 cycles. Products (354 bp) were purified and sequenced from primers 188F and 542R on an ABI3730 Genetic Analyzer by using a BigDye Terminator version 3.1 Ready Reaction Cycle Sequencing kit (Applied Biosystems, Foster City, CA, USA). Sequences were submitted to GenBank under accession nos. EF686006–13. Alignments of NP-1 gene sequences from nucleotides (nt) 2410–2602, and NP-1 predicted amino acid sequences from amino acids (aa) 1–97 were constructed by using ClustalW version 1.83 (available from www.ebi.ac.uk/tools/clustalw/index.html) and compared with HBoV prototype sequences from GenBank (DQ00495-6). Nasopharyngeal aspirates were also screened for respiratory syncytial virus (RSV) by reverse transcription–PCR (RT-PCR) and nested PCR ( 3 ) and for human metapneumovirus ( 4 ), influenza A (H1, H3), and influenza B by RT-PCR ( 5 ). Eight (3.5%) of 230 samples, collected from infants hospitalized with bronchiolitis during the 2003–2005 winter epidemic seasons, were positive for HBoV. In 5 HBoV-positive infants no other pathogens were identified, but RSV was detected in 3 (Table). The 8 HBoV-positive infants had a median age of 9.5 months, and the male:female ratio was 1:1. The median length of hospital stay was 5.5 (range 1–16) days. Table Summary of 8 infants with human bocavirus infection hospitalized with bronchiolitis, New Zealand, 2003–2005* Infant no. Date admitted Sex/ age, mo Ethnicity Attended daycare? Length of hospital stay, d Illness severity Apnea Underlying conditions/ comorbitities RSV subtype Highest temp., °C Enteritic symptoms 1 Jul 
2003 M/9 Pacific No 16 Mod – – A 40.1 Diarrhea 2 Aug 2003 F/4 Pacific No 6 Sev – – B 38.4 Diarrhea 3 Sep 2003 F/11 NZ European No 1 Mod – – – 38.1 – 4 Sep 2003 F/10 Pacific No 4 Sev – 33 weeks’ gestation – 38.3 Diarrhea 5 Aug 2004 M/8 Pacific No 2 Mod – Haemophilus influenzae conjunctivitis – 37.7 – 6 Jul 
2005 M/10 Chinese No 10 Mod – 34 weeks’ gestation, repaired esophageal atresia and tracheomalacia – 37.7 – 7 Aug 2005 F/9 Pacific No 9 Sev + 30 weeks’ gestation A 39.2 – 8 Sep 2005 M/13 NZ European Yes 5 Mod – Hydronephrosis, Pseudomonas aeruginosa urinary tract infection – 37.4 – *Temp., temperature; Mod, moderate; Sev, severe; –, absent; NZ, New Zealand;+, present. As expected, because HBoV NP-1 is highly conserved, sequence variation among New Zealand isolates and the prototype Stockholm ST-1 and ST-2 ( 1 ) NP-1 sequences was limited. Alignments of the partial NP-1 sequence (nt 2410–2602) of New Zealand isolates with those of ST-1 and ST-2 were identical, except for a G→A change at nt 176 in 2 New Zealand isolates (from infants 5 and 8 years of age), which resulted in a predicted amino acid exchange of S→N at aa 59. In addition, an A→T change at nt 274 in all 8 NZ isolates resulted in a predicted amino acid substitution of T→S at aa 92, a change that has been reported previously in Japanese isolates ( 6 ). This study reaffirms previous reports of finding HBoV in a subset of infants with bronchiolitis ( 7 ). It is also, to our knowledge, the first study of its kind in New Zealand infants, confirming wide distribution of HBoV. In the northern hemisphere, HBoV circulates primarily during the winter months, although it continues circulating until early summer, later than most other seasonal respiratory viruses ( 8 ). Therefore, this study may underestimate the percentage of New Zealand infants with bronchiolitis whose HBoV test results were positive because sample collection ceased in October (southern hemisphere spring) at the end of the bronchiolitis epidemic. The small number of HBoV-positive infants prevents conclusions concerning ethnicity, coinfection, and bronchiolitis severity. Although detection of viral nucleic acid by PCR in infants with bronchiolitis does not prove that the virus is the cause of the disease, it raises a hypothesis worthy of investigation. Further studies are required to determine the role of HBoV as a human pathogen. Although coinfection is common, HBoV detection appears to be infrequent in asymptomatic controls ( 9 ). In our study RSV was detected in 3 (37.5%) HBoV-positive samples. We may have underestimated additional coinfection because we did not test for several respiratory agents, including parainfluenza viruses, rhinoviruses, or the newly discovered coronaviruses. Finally, HBoV has recently been detected in fecal samples ( 10 ). Because 3 HBoV-positive infants had diarrhea in addition to bronchiolitis, knowing prevalence of HBoV in fecal specimens from asymptomatic New Zealand children and in those with acute gastroenteritis would be of interest.

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          Cloning of a human parvovirus by molecular screening of respiratory tract samples.

          The identification of new virus species is a key issue for the study of infectious disease but is technically very difficult. We developed a system for large-scale molecular virus screening of clinical samples based on host DNA depletion, random PCR amplification, large-scale sequencing, and bioinformatics. The technology was applied to pooled human respiratory tract samples. The first experiments detected seven human virus species without the use of any specific reagent. Among the detected viruses were one coronavirus and one parvovirus, both of which were at that time uncharacterized. The parvovirus, provisionally named human bocavirus, was in a retrospective clinical study detected in 17 additional patients and associated with lower respiratory tract infections in children. The molecular virus screening procedure provides a general culture-independent solution to the problem of detecting unknown virus species in single or pooled samples. We suggest that a systematic exploration of the viruses that infect humans, "the human virome," can be initiated.
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            Evidence of human coronavirus HKU1 and human bocavirus in Australian children

            Undiagnosed cases of respiratory tract disease suspected of an infectious aetiology peak during the winter months. Since studies applying molecular diagnostic assays usually report reductions in the number of undiagnosed cases of infectious disease compared to traditional techniques, we applied PCR assays to investigate the role of two recently described viruses, namely human coronavirus (HCoV) HKU1 and human bocavirus (HBoV), in a hospital-based paediatric population. Both viruses were found among Australia children with upper or lower respiratory tract disease during the autumn and winter of 2004, contributing to 21.1% of all microbial diagnoses, with individual incidences of 3.1% (HCoV-HKU1) and 5.6% (HBoV) among 324 specimens. HBoV was found to coincide with another virus in more than half of all instances and displayed a single genetic lineage, whilst HCoV-HKU1 was more likely to occur in the absence of another microbe and strains could be divided into two genetic lineages which we propose be termed HCoV-HKU1 type A and type B. Children under the age of 2 years were most at risk of infection by these viruses which contribute significantly to the microbial burden among patients with respiratory tract disease during the colder months.
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              Human Bocavirus Infection in Young Children in the United States: Molecular Epidemiological Profile and Clinical Characteristics of a Newly Emerging Respiratory Virus

              Abstract BackgroundHuman bocavirus (HBoV) is a newly identified human parvovirus that was originally identified in the respiratory secretions of children with respiratory tract disease. To further investigate the epidemiological profile and clinical characteristics of HBoV infection, we screened infants and children <2 years of age (hereafter referred to as “children”) for HBoV MethodsChildren for whom respiratory specimens submitted to a diagnostic laboratory tested negative for respiratory syncytial virus, parainfluenza viruses (types 1–3), influenza A and B viruses, and adenovirus, as well as asymptomatic children, underwent screening for HBoV by use of polymerase chain reaction (PCR). Respiratory specimens were obtained from the children from 1 January 2004 through 31 December 2004 ResultsTwenty-two (5.2%) of the 425 children who had a respiratory specimen submitted to the diagnostic laboratory and 0 of the 96 asymptomatic children were found to be positive for HBoV by PCR (P=.02). Fever, rhinorrhea, cough, and wheezing were observed in ⩾50% of the HBoV-positive children. Of the 17 children who had chest radiography performed, 12 (70.6%) had abnormal findings. HBoV appeared to have a seasonal distribution. Nucleotide polymorphisms were detected in the viral capsid protein (VP) 1/VP2 genes. Two distinct HBoV genotypes circulated during the study period ConclusionsHBoV is circulating in the United States and is associated with both upper and lower respiratory tract disease in infants and young children

                Author and article information

                Emerg Infect Dis
                Emerging Infect. Dis
                Emerging Infectious Diseases
                Centers for Disease Control and Prevention
                November 2007
                : 13
                : 11
                : 1797-1799
                [* ]Malaghan Institute of Medical Research, Wellington, New Zealand
                []University of Otago, Wellington, New Zealand
                Author notes
                Address for correspondence: Joanna R. Kirman, Malaghan Institute of Medical Research, PO Box 7060, Wellington South 6021, Wellington, New Zealand; email: jkirman@ 123456malaghan.org.nz
                Letters to the Editor

                Infectious disease & Microbiology
                parvoviridae,new zealand,respiratory illness,letter,bronchiolitis,diarrhea,human bocavirus


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