BackgroundHuman rhinoviruses (HRV) cause a wide spectrum of disease, ranging from a mild influenza‐like illness (ILI) to severe respiratory infection. Molecular epidemiological data are limited for HRV circulating in the Southern Hemisphere.ObjectivesTo identify the species and genotypes of HRV from clinical samples collected in Sydney, Australia, from 2006 to 2009.MethodsCombined nose and throat swabs or nasopharyngeal aspirates collected from individuals with ILI were tested for HRV using real‐time reverse‐transcriptase polymerase chain reaction (RT‐PCR). Sequencing data of 5′UTR and VP4/VP2 coding regions on RT‐PCR‐positive specimens were analysed.ResultsHuman rhinoviruses were detected by real‐time PCR in 20.9% (116/555) of samples tested. Phylogenetic analysis of 5′UTR and VP4/VP2 on HRV‐positive samples was concordant in the grouping of HRV A and B species but not HRV C species. Eighty per cent (16/20) of sequences that grouped as HRV C in the VP4/VP2 tree clustered as HRV A, alongside some previously described C strains as subspecies C/A. Discordant branching was seen within HRV A group: two sequences clustering as A in the VP4/VP2 tree branched within the C/A subspecies in the 5′UTR tree, and one sequence showed identity to different HRV A strains in the two genes. The prevalence of HRV C and C/A species was greater in paediatric compared to adult patients (47.9% vs 25.5%, P = .032).ConclusionHuman rhinoviruses are a common cause of respiratory infections, and HRV C is present in the Southern Hemisphere. Sequencing of multiple HRV regions may be necessary to determine exact phylogenetic relationships.

Background: During the early phases of the 2009 pandemic, subjects with influenza-like illness only had laboratory testing specific for the new A(H1N1)pdm09 virus.Findings: Between 25th May and 7th June 2009, during the pandemic CONTAIN phase, A(H1N1) pdm09 virus was detected using nucleic acid tests in only 56 of 1466 (3.8%) samples meeting the clinical case definition required for A(H1N1) pdm09 testing. Two hundred and fifty-five randomly selected A(H1N1) pdm09 virus-negative samples were tested for other respiratory viruses using a real-time multiplex PCR assay. Of the 255 samples tested, 113 (44.3%) had other respiratory viruses detected: rhinoviruses 63.7%, seasonal influenza A 17.6%, respiratory syncytial virus 7.9%, human metapneumovirus 5.3%, parainfluenzaviruses 4.4%, influenza B virus 4.4%, and enteroviruses 0.8%. Viral co-infections were present in 4.3% of samples.Conclusions: In the very early stages of a new pandemic, limiting testing to only the novel virus will miss other clinically important co-circulating respiratory pathogens.

Background: The aim of this study was to examine the rate of transmission of influenza and other respiratory viruses from children attending an Emergency Department to their family members in the household using active surveillance.Methods: A prospective hospital-based study was conducted over three consecutive winters (2006-2008) in children aged <1-15 years presenting with influenza-like illness (ILI). 168 children with ILI and their healthy families were recruited over three winter seasons.Results: Respiratory viruses were detected in 101 (60.8%) children with ILI; in 91/166 (54.8%) a single pathogen was detected, and in the remaining 10 children more than one virus was detected concurrently. Influenza was the most common virus detected (34/101), followed by rhinoviruses (22/101) and adenoviruses (14/101). Of influenza viruses, 21/34 were influenza A and 13/34 influenza B. Meeting the clinical definition of ILI did not differentiate between influenza and other viruses. Clinical ILI developed within one week of follow up in 12% (26/205) of the family members who were swabbed. Viral pathogens were detected in 42.3% (11/26) of the symptomatic family members. In 6/11 cases the same virus was detected in the adult and child. The lower estimate of the household risk of transmission of respiratory viruses, based on concordant proven infection in both child and adult, from a single sick child to adult household contacts is therefore 3% per week.Conclusion: This study provides quantitative, prospective data on rates of household transmission of infection from children to adults. (C) 2011 Elsevier Ltd. All rights reserved.

The more than 100 human enterovirus (HEV) serotypes can also be classified into four species, HEV-A to -D, based on phylogenetic analysis of multiple gene regions. Current molecular typing methods depend largely on reverse transcription-PCR (RT-PCR) amplification and nucleotide sequencing of the entire or 3' half of the VP1 gene. An RT-PCR-based reverse line blot (RLB) hybridization assay was developed as a rapid and efficient approach to characterize common HEVs. Twenty HEV serotypes accounted for 87.1% of all HEVs isolated at an Australian reference virology laboratory from 1979 to 2007. VP1 sequences of all known HEV prototype strains were aligned to design one sense primer and three antisense primers for RT-PCR. After sequencing of the complete VP1 genes of 37 previously serotyped examples of the commonest 20 serotypes and alignment of these VP1 sequences with GenBank sequences, four serotype-specific probes for each serotype were designed for RLB. The RT-PCR-RLB assay was then applied to 132 HEV isolates, made up of the previously sequenced 37 isolates and another 95 serotyped clinical isolates. The RT-PCR-RLB genotypes corresponded with the serotypes for 131/132 isolates; the one exception was confirmed by VP1 sequencing, and the genotype was confirmed by repeat conventional serotyping. Genotyping by RT-PCR-RLB complements traditional serotyping methods and VP1 sequencing and has the advantages of convenience, speed, and accuracy. RT-PCR-RLB allows detection of specific enteroviral serotypes or genotypes associated with HEV outbreaks and significant disease.