Hot pepper is usually attacked by various pathogens, which can cause large production loss. Molecular identification is the most commonly used method to discriminate the species of pathogens, plants, and even animals. Among the numerous DNA markers, the nuclear ribosomal DNA (nrDNA) internal transcribed spacer (ITS) region was one of the most widely used markers for pathogen species systematics. In this study, we collected 26 pepper materials from different geographical regions in South Korea, and checked and identified the pathogen species they carried based on the sequence analysis result of the nrDNA ITS region. According to the sequence analysis analogue with the existing sequence reports in the NCBI GenBank database, there were five anthracnose Colletotrichum species found in 26 materials in all, including C. acutatum, C. scovillei, C. gloeosporioides, C. nymphaeae, and C. simmondsii. Among them, C. scovillei, considered the C. acutatum species complex, combined with C. acutatum and C. gloeosporioides, were considered as the most common anthracnose pathogens in peppers, while C. nymphaeae and C. simmondsii were firstly reported to infect peppers in South Korea, although these two species had been found to infect other vegetables, fruits, and other plant species. The effective advanced detection of potential pathogens would help to direct disease prevention and treatment.