Invasive sea lamprey (Petromyzon marinus Linnaeus, 1758) are currently managed by the Great Lakes Fishery Commission in an effort to reduce pest populations below levels that cause ecological damage. One technique to improve stream population assessments could be molecular surveillance in the form of environmental DNA (eDNA) monitoring. We developed and validated four probe-based quantitative polymerase chain reaction (qPCR) assays, then used two probes (cytb, nd1) to determine whether eDNA concentration was correlated with adult and larval sea lamprey density in the lab. We found a strong positive correlation between adult sea lamprey densities of 2, 20, and 200 individuals/2000L and eDNA concentrations in tanks using both assays (cytb, nd1). For larval laboratory tank density trials, eDNA concentrations were generally near our limit of quantification and there was no significant difference in copy numbers detected between larval sea lamprey densities of 1, 5, and 25 individuals/28L. Therefore, we examined detection probability rather than concentration with laboratory tank densities. We observed a trend of increasing detection probabilities with increased larval sea lamprey density that approached significance suggesting that when DNA copy numbers are low, detection rates may be more informative in predicting varying densities of larval sea lamprey. The ability to assess sea lamprey densities from a water sample could be a powerful tool to improve traditional assessment and stream ranking techniques. Further refinement of this method in the field may make eDNA surveillance of sea lamprey a reliable part of stream assessments. Rapid eDNA analysis from many streams may help focus traditional assessment efforts, thereby improving the efficiency of invasive sea lamprey control efforts.