Within the era of smart factories, concerning the ergonomics related to production processes, the Digital Twin (DT) is the key to set up novel models for monitoring the performance of manual work activities, which are able to provide results in near real time and to support the decision-making process for improving the working conditions. This paper aims to propose a methodological framework that, by implementing a human DT, and supports the monitoring and the decision making regarding the ergonomics performances of manual production lines. A case study, carried out in a laboratory, is presented for demonstrating the applicability and the effectiveness of the proposed framework. The results show how it is possible to identify the operational issues of a manual workstation and how it is possible to propose and test improving solutions.