We developed and evaluated three qPCR assays for detecting environmental DNA from northern pike (Esox lucius). The assays target the cytochrome oxidase 1 (EluCOI), control region (EluCR), and cytochrome b (EluCytB) genes of the mitochondrial DNA. Target specificity, assessed using the fluorescence signal (at 45 cycles) to noise (at 1 cycle) ratio (S/N), showed strong amplification in northern pike (mean S/N = 2.62, 3.52, 2.69 for EluCOI, EluCR, EluCytB). The mean S/N estimates from fifteen non-esocid freshwater fishes were about 1.0, as expected for no amplification. EluCR showed evidence of amplification (mean S/N = 3.16) in muskellunge (Esox masquinongy). The sensitivity tests indicated EluCOI has a higher detection probability than EluCR and EluCytB at low (20 copies/reaction) copy number. The results favor using EluCOI although EluCytB and EluCR are viable assays for the detection of northern pike eDNA.