Biomonitoring of an invasive alien crayfish species, Procambarus clarkii (Girard, 1852), was performed from February to October 2021, along with environmental DNA analysis, at five locations including Wanju-gun, Hampyeong-gun, Naju-si, Gurye-gun, and Cheongju-si. For the investigation, an umbrella-shaped trap for adults and a hand net for young crayfish were used, and 8 to 10L of freshwater was collected for eDNA analysis. The current status and past distributional records of crayfish in Korea were analyzed along with benthic macroinvertebrates at each survey site. As a result of the investigation, a total of 122 individuals were identified, and Hampyeong-gun recorded the largest number of populations with 59 individuals (48.36%) and the highest environmental DNA (eDNA). The frequency of appearance of P. clarkii was highest in May. The ratio of females to males was 21:5, and the body size was 72.2±21.1mm for female, 80.5±15.6mm for male, and 25.3±9.8mm for young crayfish. P. clarkii introduced into Korea is mainly spreading in the southwest region and it has not been observed in Seoul where there had been a record of appearance in the past. No external symbiosis (Branchiobdellida) of P. clarkii has been identified, and P. clarkii that has appeared in Korea was presumed to be imported from Japan. There are more than eight kinds of exotic crayfish distributed in Korea, and among them, the marbled crayfish (P. virginalis) was identified as a harmful species to the ecosystem of Korea by the Ministry of Environment in 2021. The identified species of benthic macroinvertebrates inhabiting the survey area where P. clarkii has appeared were 69 belonging to 39 families, 15 orders, five classes, and three phyla. Among them, Odonata were the most abundant (16 spp.; 24.62%), followed by Coleoptera (11 spp.; 16.92%) and Hemiptera (11 spp.; 16.92%). In the survey area, one Korean endemic species (Rhoenanthus coreanus), one species (Helophorus auriculatus) classified as Near Threatened (NT) on the Korean Red List, and six species of Crustaceans appeared. In the functional feeding group, the predators appeared predominantly while in habitat oriented group, the climbers appeared to be abundant. It was confirmed that P. clarkii in the survey area prefers an area rich in aquatic vegetation with waterside vegetation, and has high resistance to turbid water quality. The omnivore invader P. clarkii is expected to maintain a competitive relationship with carnivorous benthic macroinvertebrates that are predominant in the same species, and is expected to continue to generate ecosystem disturbance along the food chains.