Infectious fungal diseases can have devastating effects on wildlife health and our ability to detect emerging fungal pathogens in the wild is considered indispensable for effective management strategies. Several fungi from the genus Nannizziopsis and Paranannizziopsis are emerging pathogens of reptiles and have been observed to cause disease in a wide range of taxa. Nannizziopsis barbatae has become a particularly important pathogen of Australian reptiles with an increasing number of herpetofauna being reported with cases of infection from across the country. Here, we report a species-specific qPCR assay for the rapid detection of N. barbatae and demonstrate its application in a wild urban population of a dragon lizard. In addition, we present the mitochondrial genome sequences and phylogenetic analysis for seven species in this group of fungi to support further research into the evolutionary ecology of these emerging pathogens.