Public health laboratories require rapid diagnosis of dengue outbreaks for application of measures such as vector control. We have developed a rapid single fluorogenic probe-based polymerase chain reaction assay for the detection of all four dengue serotypes (FUDRT-PCR). The method employs primers and probe that are complementary to the evolutionarily conserved 3' untranslated region of the dengue genome. The assay detected viral RNA of strains of all four dengue serotypes but not of the flaviviruses Japanese encephalitis virus, Murray Valley encephalitis virus, Kunjin, Stratford, West Nile, Alfuy or Yellow fever. When compared to an existing nested-PCR assay for the detection of dengue on clinical samples, FUDRT-PCR detected dengue 1 (100%, n=14), dengue 2 (85%, n=13), dengue 3 (64%, n=14) and dengue 4 (100%, n=3) with the indicated sensitivities. FUDRT-PCR enables diagnosis of acute dengue infection in four hours from sample receipt. In addition, a single-test procedure should result in a reduction in the number of tests performed with considerable cost savings for diagnostic laboratories.

Molecular epidemiologic analysis shows that travelers returning from Asia are the greatest source of risk.

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Since the establishment of West Nile virus (WNV) into the United States, concern has arisen that this virus may also pose a serious threat to Australian biosecurity. The vector competence of 19 Australian mosquito species for a North American strain of WNV was evaluated. Mosquitoes collected from Cairns, Brisbane, and Sydney were exposed to blood containing 10(4.0+/-0.3) cell culture infectious dose(50)/mosquito WNV that was isolated from a crow during the 1999 New York outbreak. Mosquitoes were tested 12-15 days later to determine their infection, dissemination, and transmission rates. A number of Culex spp. demonstrated a high vector competence for this virus, with some populations of Culex annulirostris, the primary Australian Kunjin virus vector, displaying transmission rates up to 84%. Similarly, Cx. quinquefasciatus and Cx. gelidus were highly competent, with infection and transmission rates of >80% and >50%, respectively. Common Aedes spp., including Aedes notoscriptus, Ae. vigilax, and Ae. procax, were moderately susceptible, and some Verrallina spp. and Coquillettidia spp. were relatively refractory to infection. Thus, Australia possesses a number of competent mosquito species that could facilitate local transmission of WNV, should it be introduced.

In response to an incursion of Japanese encephalitis virus (JEV) on Cape York Peninsula, Australia, in 2005, 23,144 Culex mosquitoes were processed for virus detection. A single isolate of JEV was obtained from a pool of Culex sitiens subgroup mosquitoes. This is the first reported mosquito isolate of JEV from the Australian mainland.