Background: To explore the reasonable orientation of talent training for medical laboratory technology (MLT) at the vocational undergraduate level, this study has conducted a thorough survey and analysis of the talent market demand in multiple aspects, including the total MLT talent market demand, talent demand changes and expectations, and changes in industry development. Methods: Based on a literature search, this study collected statistics on the number of MLT professionals in China over the past decade. Questionnaires were distributed to 128 employers in 14 provinces nationwide. A survey was conducted on the overall industry development and the requirements of enterprises for the abilities of talent on the market, and 26 doctors and nurses with senior technical titles were interviewed. Another survey was carried out among 17 higher vocational colleges and 8 general undergraduate universities nationwide, focusing on vocational abilities that are highly regarded by teachers. Results: As reflected by the results of this study, the market requires a higher level of educational background for talent, and self-directed learning ability is rated as the most essential vocational ability by enterprises. Moreover, the ability to use information technology and the communication and expression ability are necessary personal abilities in the opinion of enterprises. However, the vocational abilities specially developed by teachers are not always the same as those required by enterprises. The results of this study show that the latest progress of information technology in the industry must be integrated into talent training, and priority should be given to developing innovation ability, self-directed learning ability and interpersonal communication ability to develop talent that precisely meets market demand. Conclusion: Based on a thorough survey and analysis of market demand, this study recommends upgrading traditional teaching thought, focusing on specific market demand, accurately determining talent training and ensuring that talent can better meet market demand.