Sir Albert Sakzewski Virus Research Center

Australia

Sir Albert Sakzewski Virus Research Center

Australia

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Preston F.M.,University of Queensland | Preston F.M.,Sir Albert Sakzewski Virus Research Center | Straub C.P.,University of Queensland | Straub C.P.,Sir Albert Sakzewski Virus Research Center | And 4 more authors.
Virology Journal | Year: 2012

Background: Human metapneumovirus (hMPV) is a significant viral respiratory pathogen of infants and children, the elderly and immunocompromised individuals. Disease associated with hMPV infection resembles that of human respiratory syncytial virus (RSV) and includes bronchiolitis and pneumonia. The glycosylated G attachment protein of hMPV is required for viral entry in vivo and has also been identified as an inhibitor of innate immune responses. Findings: We designed and validated two siRNA molecules against the G gene using A549 cells and demonstrated consistent 88-92% knock-down for one siRNA molecule, which was used in subsequent experiments. Significant reduction of G mRNA in A549 cells infected with hMPV did not result in a reduction in viral growth, nor did it significantly increase the production of type I interferon (α/β) in response to infection. However, there was a moderate increase in IFN-β mRNA expression in response to infection in siG-transfected cells compared to untransfected and si-mismatch-transfected cells. Expression of G by recombinant adenovirus did not affect type I IFN expression. Conclusion: G has been previously described as a type I interferon antagonist, although our findings suggest it may not be a significant antagonist. © 2012 Preston et al.; licensee BioMed Central Ltd.


Goire N.,University of Queensland | Goire N.,Sir Albert Sakzewski Virus Research Center | Lahra M.M.,The Prince of Wales Hospital | Ohnishi M.,Japan National Institute of Infectious Diseases | And 10 more authors.
Eurosurveillance | Year: 2013

Emergence and spread of Neisseria gonorrhoeae resistant to extended spectrum cephalosporins is a major problem threatening treatment of gonorrhoea and is further highlighted by the recent report of a second ceftriaxone-resistant N. gonorrhoeae strain (F89) in Europe, initially observed in France and subsequently identified in Spain. N. gonorrhoeae antimicrobial resistance (AMR) surveillance has acquired new importance and molecular tools have the potential to enhance bacterial culture-based methods. In this study, we established a polymerase chain reaction (PCR) protocol for direct detection of the F89 strain. A key component of this screening protocol was the development of a hybridisation probe-based melting curve analysis assay (mosaic501-hybPCR) to detect the presence of an A501P substitution on the N. gonorrhoeae mosaic penicillin binding protein 2 (PBP2) sequence, an important characteristic of the F89 strain. The mosaic501-hybPCR was evaluated using plasmid-derived positive controls (n=3) and characterised gonococcal (n=33) and non-gonococcal (n=58) isolates. The protocol was then applied to 159 clinical specimens from Sydney, Australia, collected during the first half of the year 2012 that were N. gonorrhoeae PCR-positive. Overall, the results indicate that the PCR-based protocol is suitable for direct detection of the N. gonorrhoeae F89 strain in non-cultured clinical samples. It therefore provides an additional tool to aid investigations into the potential spread of F89 strain throughout Europe and elsewhere.


Wei T.,Queensland Institute of Medical Research Berghofer | Li D.,Queensland Institute of Medical Research Berghofer | Marcial D.,Queensland Institute of Medical Research Berghofer | Marcial D.,University of Queensland | And 12 more authors.
PLoS ONE | Year: 2014

The eukaryotic translation factor eEF1A assists replication of many RNA viruses by various mechanisms. Here we show that down-regulation of eEF1A restricts the expression of viral genomic RNA and the release of infectious virus, demonstrating a biological requirement for eEF1A in the respiratory syncytial virus (RSV) life cycle. The key proteins in the replicase/transcriptase complex of RSV; the nucleocapsid (N) protein, phosphoprotein (P) and matrix (M) protein, all associate with eEF1A in RSV infected cells, although N is the strongest binding partner. Using individually expressed proteins, N, but not P or M bound to eEF1A. This study demonstrates a novel interaction between eEF1A and the RSV replication complex, through binding to N protein, to facilitate genomic RNA synthesis and virus production. © 2014 Wei et al.


Dang X.,Beth Israel Deaconess Medical Center | Bialasiewicz S.,Sir Albert Sakzewski Virus Research Center | Bialasiewicz S.,Queensland Childrens Medical Research Institute | Nissen M.D.,Sir Albert Sakzewski Virus Research Center | And 5 more authors.
PLoS ONE | Year: 2011

Conflicting prevalence of newly identified KI(KIPyV), WU(WUPyV) and Merkel Cell Carcinoma(MCPyV) polyomaviruses have been reported in progressive multifocal leukoencephalopathy(PML) patient samples, ranging from 0 to 14.3%. We analyzed the prevalence of these polyomaviruses in cerebrospinal fluid(CSF), peripheral blood mononuclear cells(PBMC), and bone marrow samples from PML patients, immunosuppressed individuals with or without HIV, and multiple sclerosis(MS) patients. Distinct PCR tests for KIPyV, WUPyV and MCPyV DNA performed in two independent laboratories detected low levels of MCPyV DNA only in 1/269 samples. The infrequent detections of these viruses in multiple samples from immunosuppressed individuals including those with PML suggest that their reactivation mechanisms may be different from that of JC polyomavirus (JCPyV) and that they do not play a role in the pathogenesis of PML. © 2011 Dang et al.


Bialasiewicz S.,University of Queensland | Bialasiewicz S.,The Royal Childrens Hospital | Bialasiewicz S.,Sir Albert Sakzewski Virus Research Center | McVernon J.,Murdoch Childrens Research Institute | And 8 more authors.
BMC Infectious Diseases | Year: 2014

Background: Human Parainfluenza viruses are a common cause of both upper and lower respiratory tract infections, particularly in children. Of the four Parainfluenza virus serotypes, Parainfluenza 4 is least well characterised from both the clinical, epidemiological and genetic perspectives. Methods: Flocked nose or throat swabs from a previous study investigating viral prevalence in community-based adults suffering from influenza like illness were used as the basis for this study. Samples in which no virus was detected using a 16 viral respiratory pathogen real-time PCR panel were barcoded and pyrosequenced using the Roche 454 GS FLX Titanium chemistry. The sequences were analysed using the VirusHunter bioinformatic pipeline. Sanger sequencing was used to complete the detected Parainfluenza 4 coding region. Results: A variant Parainfluenza 4 subtype b strain (QLD-01) was discovered in an otherwise healthy adult who presented with influenza like illness. Strain QLD-01 shared genomic similarities with both a and b subtypes. The extent of divergence of this genome from the 5 available whole Parainfluenza 4 genomes impacted the predicted binding efficiencies of the majority of published Parainfluenza 4 PCR assays. Conclusions: These findings further support a possible role for Parainfluenza 4 in the aetiology of adult respiratory disease within the community setting, and highlight the caution needed to be used in designing PCR assays from limited sequence information or in using proprietary commercial PCR assays. © 2014 Bialasiewicz et al.; licensee BioMed Central Ltd.


Bialasiewicz S.,Sir Albert Sakzewski Virus Research Center | MacKay I.M.,Sir Albert Sakzewski Virus Research Center | Whiley D.M.,Sir Albert Sakzewski Virus Research Center | Sloots T.P.,Sir Albert Sakzewski Virus Research Center
Methods in Molecular Biology | Year: 2012

Etiologic agents of genital ulcer disease include herpes simplex 1 and 2, Treponema pallidum pallidum, Haemophilus ducreyi, and Klebsiella granulomatis. The advent of PCR has allowed for more rapid and sensitive detection of microbial pathogens. In this protocol, we describe the simultaneous detection of these five pathogens and an internal control using a single-tube multiplex PCR and colorimetric enzyme-linked amplicon hybridization assay. © 2012 Springer Science+Business Media New York.


Goire N.,University of Queensland | Sloots T.P.,Sir Albert Sakzewski Virus Research Center | Nissen M.D.,University of Queensland | Whiley D.M.,Sir Albert Sakzewski Virus Research Center
Methods in Molecular Biology | Year: 2012

Gonorrhoea is no longer an easily treatable ailment but rather is now a challenging disease in terms of antimicrobial resistance (AMR) with treatment options rapidly diminishing. The causative agent of gonorrhoea, Neisseria gonorrhoeae, has managed to develop resistance to almost every single drug used against it with the sole exception of extended spectrum cephalosporins. The situation is further exacerbated by the fact that not only are the rates of gonococcal infections on a steady rise globally, but tracking AMR is being undermined by the growing popularity of molecular methods at the expense of traditional bacterial culture in diagnostic laboratories. Recently, concerns have been raised over the emergence of a multi-resistant gonococci and the potential for untreatable gonorrhoea. Maintaining optimal epidemiological surveillance of gonococcal AMR remains an important aspect of gonorrhoea control. The development of molecular tools for tracking AMR in N. gonorrhoeae has the potential to further enhance such surveillance. In this chapter, we discuss nucleic acid amplification-based detection of AMR in gonorrhoea with a particular emphasis on chromosomal-mediated resistance to beta-lactam antibiotics. © 2012 Springer Science+Business Media New York.


Lambert S.B.,Sir Albert Sakzewski Virus Research Center | Lambert S.B.,University of Queensland | Chuk L.-M.R.,Sir Albert Sakzewski Virus Research Center | Chuk L.-M.R.,University of Queensland | And 21 more authors.
Influenza and other Respiratory Viruses | Year: 2013

Objective: To evaluate the safety of CSL's split-virion inactivated trivalent 2009 Southern Hemisphere formulation influenza vaccine (TIV) in children. Methods: We enrolled 1992 healthy children into three groups: Cohorts A, ≥6 months to <3 years; B, ≥3 years to <9 years; and C, ≥9 years to <18 years. Children received one or two doses of 0·25 ml (22·5 μg haemagglutinin) or 0·5 ml (45 μg) TIV, depending on age and prior vaccination history. We collected post-vaccination solicited adverse event (AE) data (days 0-6), including fever (temperature: ≥37·5°C axilla, ≥38·0°C oral), unsolicited AEs (days 0-29) and serious AEs (SAEs) and new-onset chronic illnesses (NOCIs; to day 180 after last vaccination). Results: At least one solicited AE was reported by 80%/78%/78% of children in Cohorts A, B and C, respectively. Systemic AEs were more common among Cohort A (72% of participants), and local AEs were more common among Cohort C (71% of participants). Fever was more common in younger cohorts, in influenza vaccine-naïve children (29% of Cohort A receiving their first dose), and following first compared with second doses. Severe fever following a first dose prevented 20 participants receiving their second scheduled vaccine dose. A 7-month-old participant had a single uncomplicated febrile convulsion on the day of vaccination. Conclusions: Nearly 80% of subjects reported at least one solicited AE following immunization. Fever prevalence was highest in vaccine-naïve Cohort A participants, similar to other paediatric studies using CSL vaccine. Further research to understand fever-related AEs in children following CSL's TIV is recommended. © 2013 John Wiley & Sons Ltd.


PubMed | Sir Albert Sakzewski Virus Research Center
Type: Journal Article | Journal: Journal of virology | Year: 2010

Exploration of the genetic diversity of WU polyomavirus (WUV) has been limited in terms of the specimen numbers and particularly the sizes of the genomic fragments analyzed. Using whole-genome sequencing of 48 WUV strains collected in four continents over a 5-year period and 16 publicly available whole-genome sequences, we identified three main WUV clades and five subtypes, provisionally termed Ia, Ib, Ic, II, IIIa, and IIIb. Overall nucleotide variation was low (0 to 1.2%). The discriminatory power of the previous VP2 fragment typing method was found to be limited, and a new, larger genotyping region within the VP2/1 interface was proposed.

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