On the Home Front: Specialised Reference Testing for Dengue in the Australasian Region

Abstract

Reference laboratories are vital for disease control and interpreting the complexities and impact of emerging pathogens. The role of these centralized facilities extends beyond routine screening capabilities to provide rapid, specific, and accurate diagnoses, advanced data analysis, consultation services, and sophisticated disease surveillance and monitoring. Within the Australasian region, the Public Health Virology Laboratory (PHV), Forensic and Scientific Services, Department of Health, Queensland Government, Australia, and the Institute of Environmental Science and Research Limited (ESR), New Zealand (NZ) perform specialised reference testing and surveillance for dengue viruses (DENVs) and other emerging arthropod-borne viruses (arboviruses), including chikungunya virus (CHIKV) and Zika virus (ZIKV). With a focus on DENV, we review the reference testing performed by PHV (2005 to 2017) and ESR (2008 to 2017). We also describe how the evolution and expansion of reference-based methodologies and the adoption of new technologies have provided the critical elements of preparedness and early detection that complement frontline public health control efforts and limit the spread of arboviruses within Australasia.

A Case of Japanese Encephalitis with a Fatal Outcome in an Australian Who Traveled from Bali in 2019

“…The CSF sample also yielded a JEV isolate following one passage in Aedes albopictus mosquito C6/36 cells ( Figure 2). Specific patient anti-JEV IgM antibodies were also detected by FL-MIA in a serum sample collected on day six after onset of symptoms, and an anti-flavivirus IgG antibody seroconversion determined via an in-house-developed enzyme-linked immunosorbent assay (ELISA) [4] was confirmed in a serum sample collected on day 12 of his illness. The CSF collected on day four of illness was submitted for microbiological investigations.…”

“…Specific patient anti-JEV IgM antibodies were also detected by FL-MIA in a serum sample collected on day six after onset of symptoms, and an anti-flavivirus IgG antibody seroconversion determined via an in-house-developed enzyme-linked immunosorbent assay (ELISA) [ 4 ] was confirmed in a serum sample collected on day 12 of his illness.…”

“…In vitro culture of Japanese encephalitis virus (JEV) from the cerebrospinal fluid (CSF) sample in Aedes albopictus C6/36 cells. Cell culture monolayers were analysed by immunofluorescent antibody assay (IFA)[4] using an anti-JEV monoclonal antibody, 6B4A-10 (Catalogue number MAB8743, Merck, Sigma-Aldrich, Australia). (A) CSF-inoculated, IFA-positive (green stain), JEVinfected C6/36 cells and (B) uninfected, IFA-negative C6/36 cells.Specific patient anti-JEV IgM antibodies were also detected by FL-MIA in a serum sample collected on day six after onset of symptoms, and an anti-flavivirus IgG antibody seroconversion determined via an in-house-developed enzyme-linked immunosorbent assay (ELISA)[4] was confirmed in a serum sample collected on day 12 of his illness.…”

“…In vitro culture of Japanese encephalitis virus (JEV) from the cerebrospinal fluid (CSF) sample in Aedes albopictus C6/36 cells. Cell culture monolayers were analysed by immunofluorescent antibody assay (IFA)[4] using an anti-JEV monoclonal antibody, 6B4A-10 (Catalogue number MAB8743, Merck, Sigma-Aldrich, Australia). (A) CSF-inoculated, IFA-positive (green stain), JEV-infected C6/36 cells and (B) uninfected, IFA-negative C6/36 cells.…”

Erratum: Pyke, A.T. et al. On the Home Front: Specialized Reference Testing for Dengue in the Australasian Region. Trop. Med. Infect. Dis. 2018, 3, 75

“…The authors wish to make the following correction to Table 1 of this paper [1]: The DENV 1-4 DU5 MGB2017 real-time polymerase chain reaction (RT-rtPCR) assay should state that it is designed in the Capsid peptide coding region under the genome region column, and not the 3'UTR.…”