Fig. 2

HRM real-time qPCR analysis for the identification of arthropod-borne filarioids in Australia dogs. Normalized melting curves (a) and difference melting curves (b). Curves include positive controls of Acanthocheilonema reconditum (green) and Dirofilaria immitis (red, blue). To generate normalised melting curves and difference melting curves, pre- (70–71 °C) and post-melt (76–77 °C) normalisation regions were set to define the main temperature boundaries of the normalised and difference plots. The Precision Melt Software v. 1.2 (BioRad) then assigned each sample to a cluster (Clusters 1–3) using default settings. Single dog sample was assigned to A. reconditum (light green; note the ambiguous melting profile in differential melting graph b), however DNA sequencing matched D. immitis 12S rRNA gene reference sequence (FN391554)