Leptospirosis is a zoonotic bacterial infection caused by Leptospira spp., a spirochaete with global distribution and host-species endemicity among Australian native fauna and domestic animals. This disease poses a threat to public health, with clinical symptoms ranging from mild, self-limiting fever to potentially life-threatening complications including multi-organ failure and pulmonary haemorrhage. Leptospirosis tends to occur more frequently in humid, tropical environments where the bacteria are excreted into the environment through animal urine. As Australian climatic and population dynamics evolve in contemporary times and into the future, opportunities for human exposure to the sources of Leptospiral infection, which include zoonotic contact with host mammalian species and contaminated environments, will potentially increase. Population growth requires the expansion of urban infrastructure into host species habitats and intensification of farming practices to cater the food requirements of a larger population. Furthermore, the frequency and severity of natural disasters affecting Australia, including flooding events, are likely to become more frequent due to climate change. Occupational exposure in the agriculture industry, contact with infected animals and exposure to flood waters all comprise known risk factors for the development of Leptospirosis. Therefore, the risk posed to human health by Leptospiral infection is hypothesised to potentially increase in coming years and indeed, current data suggests that incidence of infection has increased in Queensland in the last 15 years. Currently the methods available for laboratory diagnosis are largely reliant on serology via the gold standard Microscopic Agglutination Test, attempts at culture and isolation using specialised culture media and Nucleic Acid Analysis utilising the Lipl32 gene target. Each of these methods is highly sensitive and specific but not without limitations, namely the limitations posed by the timing of clinical suspicion and narrow, specified windows for appropriate specimen collection within the Leptospirosis disease progression paradigm. This presents a significant challenge for Leptospirosis diagnostics but also for epidemiological monitoring. As we look to the future of Leptospiral infection diagnosis, disease management and surveillance, new prospects for laboratory diagnostic and typing techniques should be explored to meet these challenges. Recent progress in genomic sequencing of the reference culture collection will facilitate this.