We all get a cold and the sniffles now and then, but are respiratory infections as harmless as they seem? The worldwide outbreak of H1N1’09 virus (‘swine flu’) has shown that there is still much to learn about the transmission of respiratory pathogens. We have developed a number of multiplex PCRs and Real-time PCRs for detection of multiple viral and bacterial respiratory pathogens in order to learn more about the transmission of respiratory pathogens and their importance in chronic respiratory diseases such as asthma and cystic fibrosis. We are also interested in the evolution of influenza, and how it causes epidemics and pandemics. Our laboratory played an important role in collaboration with the South East Area Laboratory Service (SEALS) Virology Diagnostic Laboratory in detecting the first Sydney cases of the 2009 Swine Flu outbreak.
In 2018 we are particularly interested in:
Cystic Fibrosis and Respiratory Viruses
Cystic Fibrosis (CF) is the most common life-threatening, genetic disorder amongst Australians. In Australia, one baby with CF is born every four days. In collaboration with the two largest children’s hospitals in Sydney (The Children’s Hospital at Westmead and The Sydney Children’s Hospital Randwick), we have studied large cohorts of children suffering from CF for respiratory viral and bacterial infections, and the effect these infections have on their condition. In particular, we hope to learn more about the role of bacterial and viral infections on CF progression and on the sudden worsening of symptoms. We are exploring novel methods for better monitoring these infections.
Asthma and Respiratory Viruses
Asthma exacerbations are frequently triggered by viral respiratory infections. The types of viruses identified include HRV, respiratory syncytial virus (RSV), influenza, coronaviruses, parainfluenza, enteroviruses, human metapneumovirus (HMPV) and bocavirus. We have recently completed an NH&MRC grant to study children at risk of an asthma exacerbation and monitor them longitudinally to examine the respiratory viruses they become infected with, and how this affects their asthma. The publications related to this study can be found below.
Point-of-care testing for respiratory viruses
Current diagnostic testing for respiratory viruses is by polymerase chain reaction (PCR), which is sensitive, although typical turn-around-time (TAT) to diagnosis is approximately one day. Utilising at patient bedside, rapid “point-of-care tests” (POCT) tests in the emergency department can result in the reduced length of hospital stay and increased antiviral prescription patients with influenza. Recent developments of new version POCT have a TAT of around 30 min, and are highly sensitive and specific for influenza and other respiratory viruses. We are using these POCT in different cohorts to measure improvement in patient outcomes.
Transmission of Respiratory Viruses
Viruses can be spread by both contact and airborne routes (Gralton et al, 2010 ePub). We have previously demonstrated viral RNA in expelled particles produced on coughing and breathing (Stelzer-Braid et al., 2009). Transmission via other modes is less well described in the literature. Of particular interest to us is transmission via contact with bed linen and furniture, which is a recognised route of transmission of respiratory viruses in humans and animals. In this study, based in childcare centres, we are investigating whether contamination of personal clothing, an unusual fomite, has the potential to propagate the spread of respiratory pathogens by environmental and occupational exposure.
Novel sampling methods
In collaboration with A/Prof Euan Tovey from the Woolcock Institute of Medical Research at the University of Sydney, we have developed novel methods for collection and measurement of respiratory viruses. We have developed a mask containing a special filter that non-invasively samples viruses exhaled during coughing and breathing. We have successfully recovered viral RNA and culturable virus off this filter and have published these results (Stelzer-Braid et al, 2009). We are currently refining the design. Our methods have real potential in non-invasive sampling, which would be useful for all kinds of chronic respiratory and infectious diseases.
Fidelity and Evolution of Influenza
Influenza virus evolves continuously through mutations presumed to result from evolutionary pressure driving viral replication. This study examines the relationship between the genetic evolution of a new antigenic variant of influenza virus in a human population using sequence data and sequencing plaque assay, and revealed a gradual decrease in the rate of genetic evolution of the new antigenic variant. It is possible the replication fidelity of the influenza polymerase contributes to the evolution of influenza viruses. We extend this to examine other methods of assessing replication fidelity of influenza viruses, and are currently using these methods to assess the replication fidelity of the recent 2009 pandemic A/H1N1 influenza viruses, which would be important in pandemic surveillance and vaccine prediction.
Most Recent Publications
Dyda, A., Stelzer-Braid, S., Adam, D., Chughtai, A.A., MacIntyre, C.R. (2018) The association between acute flaccid myelitis and Enterovirus D68 (EV- D68) – what is the evidence for causation? Eurosurveillance, 23(3), 18/Jan/2018. IF: 5.722.
Stelzer-Braid S., Liu N., Doumit M., D'Cunha R., Belessis Y., Jaffe A., Rawlinson W.D. (2017) Association of rhinovirus with exacerbations in young children affected by cystic fibrosis: Preliminary data. Journal of Medical Virology, 89(8): 1494-1497.
Jensen, T.O., Stelzer-Braid, S., Willenborg, C., Cheung, C., Andresen, D., Rawlinson, W.D. and Clezy K. (2016) Outbreak of Respiratory Syncytial Virus (RSV) infection in immunocompromised adults on a hematology ward. Journal of Medical Virology, accepted 14 March 2016. DOI 10.1002/jmv.24521
Doumit, M., Belessis, Y., Stelzer-Braid, S., Mallitt, K., Rawlinson W.D., and Jaffe A. (2015) Diagnostic accuracy and distress associated with oropharyngeal suction in cystic fibrosis. Journal of Cystic Fibrosis, accepted Sep 17 2015. Doi: 10.1016/j.jcf.2015.09.001
Stelzer-Braid S., Tovey E.R., Willenborg C.M., Toelle B.G., Ampon R., Garden F.L., Oliver B.G., Strachan R., Belessis Y., Jaffe A., Reddel H.K., Crisafulli D., Marks G.B. and Rawlinson W.D. (2015) Absence of back to school peaks in human rhinovirus detections and respiratory symptoms in a cohort of children with asthma. Journal of Medical Virology, doi: 10.1002/jmv.24371.
Homaira N, Sheils J, Stelzer-Braid S, Lui K, Oie JL, Snelling T, Jaffe A and Rawlinson W. (2015) Respiratory syncytial virus is present in the neonatal intensive care unit. Journal of Medical Virology, http://dx.doi.org/10.1002/jmv.24325
Tovey E.R., Stelzer-Braid S., Toelle B.G., Oliver B.G., Reddel H.K., Willenborg, C.M., Belessis, Y. C., Garden, F.L., Jaffe, A., Strachan, R., Eyles, D., Rawlinson, W.D. and Marks, G.B. (2014) Rhinoviruses significantly affect day-to-day respiratory symptoms of children with asthma. Journal of Allergy and Clinical Immunology, http://dx.doi.org/10.1016/j.jaci.2014.10.020,
Willenborg C; Stelzer-Braid SJ (2013) Sneezing leads to wheezing: microorganisms important in asthma, Microbiology Australia, vol. 34, no. 3, http://microbiology.publish.csiro.au/?paper=MA13042
Wong KKY; Rockman S; Ong C; Bull R; Stelzer-Braid S; Rawlinson W (2013) Comparison of influenza virus replication fidelity in vitro using selection pressure with monoclonal antibodies, Journal of Medical Virology, vol. 85, no. 6, pp. 1090 - 1094, http://dx.doi.org/10.1002/jmv.23532
Stelzer-Braid S., Johal H., Skilbeck K., Steller A., Alsubie H., Tovey E., Van Asperen P., McKay K. and Rawlinson W.D. (2012) Detection of viral and bacterial respiratory pathogens in patients with cystic fibrosis. Journal of Virological Methods 186:109-112. http://dx.doi.org/10.1016/j.jviromet.2012.08.008
Rawlinson WD, Iwasenko JM, Robertson P, Taylor PC (2011) Accuracy of Influenza diagnosis. Letter to the editor. Infection Control and Hospital Epidemiology, in press
Tovey E, Rawlinson WD (2011) A modern miasma hypothesis and back-to-school asthma exacerbations. Medical Hypothesis, in press
Wong K, Bull R, Rockman S, Scott G, Stelzer-Braid S, Rawlinson WD (2011) Correlation of polymerase replication fidelity with genetic evolution of influenza A/Fujian/411/02(H3N2) viruses. Journal of Medical Virology 83(3):510-516
Gralton J, Tovey E, McLaws M-L, Rawlinson W (2011) The role of particle size in aerosolised pathogen transmission: A review. Journal of Infection 60(1):1-13
Iwasenko JM, Cretikos M, Paterson DL, GibbR, Webb SA, Smith DW, Blyth CC, Dwyer DE, Shi JQ, Robertson P, Rawlinson WD (2010) Enhanced diagnosis of pandemic (H1N1) 2009 influenza infection using molecular and serological testing in intensive care unit patients with suspected influenza. Clinical Infectious Diseases 51(1):70-72.
Blyth CC, Foo H, van Hal SJ, Hurt AC, Barr IG, McPhie K, Armstrong PK, Rawlinson WD, Sheppeard V, Conaty S, Staff M, Dwyer DE, World Youth Day I (2010) Influenza outbreaks during World Youth Day 2008 mass gathering. Emerging Infectious Diseases 16(5):809-15
Dalton CB, Cretikos MA, Durrheim DN, Seppelt IM, Rawlinson WD, Dwyer DE (2010) Comparison of adult patients hospitalised with pandemic (H1N1) 2009 influenza and seasonal influenza during the "PROTECT" phase of the pandemic response. Medical Journal of Australia 192(6):357-8
Foo H, Blyth CC, van Hal S, McPhie K, Ratnamohan M, Fennell M, BaAlawi F, Dwyer D, Rawlinson W, Adamson S, Armstrong P, Dwyer DE (2009) Laboratory test performance in young adults during influenza outbreaks at world youth day 2008. Journal of Clinical Virology 46:384-386.
Hurt AC, Ernest J, Deng Y-M, Iannello P, Besselaar T, Birch C, Buchy P, Chittaganpitch M, Chiu S-C, Dwyer D, Guigon A, Harrower B, Ip PK, Kok T, Lin C, McPhee K, Mohd A, Olveda R, Panayotou T, Scott L, Smith D, Smith L, Rawlinson W, Komadina N, Shaw R, Kelso A, Barr IG (2009) Emergence and spread of oseltamivir-resistant A(H1N1) influenza viruses in Oceania, South East Asia and South Africa. Antiviral Research 83:90-93
Stelzer-Braid S, Oliver BG, Blazey AJ, Argent E, Newsome TP, Rawlinson WD, Tovey ER (2009) Exhalation of respiratory viruses by breathing, coughing and talking. Journal of Medical Virology 81(9): 1674-1679
Huynh KB, Oliver BG, Stelzer S, Rawlinson WD, Tovey ER (2008) A new method for sampling and detection of exhaled respiratory virus aerosols. Clinical Infectious Diseases 46:93–95
Lynch GW, Selleck PW, Axell AM, Downton T, Kapitza N, Boehm I, Dyer W, Wang Y-FY, Stelzer-Braid S, Rawlinson W, Sullivan JS (2008) Cross-Reactive Anti-Avian H5N1 Influenza Neutralizing Antibodies in a Normal ‘Exposure-Naïve’ Australian Blood Donor Population. Open Immunology Journal 1:13-19
Stelzer-Braid S, Escott R, Baleriola C, Kirkland P, Robertson P, Catton M, Rawlinson WD (2008) Proficiency of nucleic acid tests for avian influenza viruses, Australasia. Emerging Infectious Diseases 14:1126-1128
Stelzer-Braid S, Wong B, Robertson P, Lynch G, Laurie K, Shaw R, Barr I, Selleck PW, Baleriola C, Escott R, Katsoulotos G, Rawlinson WD (2008) A commercial ELISA detects high levels of human H5 antibody but cross-reacts with influenza A antibodies. Journal of Clinical Virology 43(2):241-243
Tozer S, Whiley DM, Lum G, Rawlinson WD, Sammels LM, Sloots TP, Nissen MD (2008) Identification of Australian human respiratory syncytial virus strains containing a 60-nucleotide duplication within the G glycoprotein gene. Pathology 40(6):632-635