The core of “Double First Class” construction is to cultivate first-class talents, and courses are the core element of talent training. The construction of application-oriented demonstration courses is an important measure for local colleges and universities to cultivate applied talents, and its construction level directly affects the quality of personnel training. This article takes the “Clinical Microbiology Testing” course of Chengdu Medical College as an example, and introduces the reform and practice of the course around the course objectives, course content, teaching methods, and course assessment. These reforms pay more attention to the cultivation of professional knowledge, practical ability, innovation ability, and humanistic literacy in teaching, highlighting the training goal of high-quality clinical microbiological testing applied talents. Through years of reform and development, this course has formed a distinctive medical curriculum system, which can provide a reference for the construction of application-oriented demonstration courses in local medical colleges.