Environmental DNA (eDNA) detection method is considered as a new survey tool for the wildlife aquatic animals in recent years. To further investigate the Hong Kong newt (Paramesotriton hongkongensis) population distribution in the wild, we thus aimed to design a set of specific primers and TaqMan probe and verify the feasibility of this eDNA method for water samples. The mitochondrial cytochrome b genes of 11 Paramesotriton species were sequenced, and then the primers and TaqMan probe were designed using Primer Express Software 3.0. The primers and TaqMan probe specificity were verified by comparing to the NCBI GenBank and testing with the annealing temperature gradient. The qPCR amplification efficiency was optimized by testing different concentrations of primers and TaqMan probe. Then, eDNA was detected in the water samples in aquarium tanks with different numbers of Hong Kong newt to assess the sensitivity of the established qPCR method. Also, the decay rate of Hong Kong newt eDNA was determined to estimate its lasting time in water. The results showed that the designed primers and TaqMan probe only amplified eDNA of Hong Kong newt but not the other 10 closely related newts (Table 3 and 4). The optimized qPCR achieved the efficiency of 93.9 % and the limit of detection down to 10 DNA copies (Fig. 2 and Table 5). The established qPCR method was sensitive enough to detect 13.56 ± 3.35 DNA copies/ml in the aquarium tank with 1 Hong Kong newt after housing for 24 h (Fig. 3). In addition, the decay rate experiment demonstrated that the use of 0.45 μm pore size filter membrane was effective to detect and monitor the Hong Kong newt eDNA in 15 d (Fig. 6). In this study, an eDNA detection method was successfully designed and established to detect the presence or absence of Hong Kong newt in water environments, which can be potentially utilized in the field inspection.