This report is a summary of the 4th Conference on NGS for Adventitious Virus Detection, which took place on December 4-5, 2024, in Frankfurt, Germany, and was sponsored by the International Alliance for Biological Standardization (IABS), and co-chaired by the U.S. Food and Drug Administration (FDA) and the European Directorate for the Quality of Medicines & HealthCare (EDQM). The increased interest in using NGS for adventitious virus detection follows its recent introduction in the ICH Q5A (R2) guideline and the new EDQM/European Pharmacopoeia general chapter 2.6.41. Key conference objectives included evaluating NGS validation and implementation, addressing regional challenges, and discussing regulatory acceptance as an alternative method to the conventional assays. The conference fostered networking between early and established NGS users and emphasized the Advanced Virus Detection Technologies Working Group as a key learning hub for NGS applications. Discussions focused on method validation requirements and the need for defining a specific limit of detection. Participants shared updates on scientific developments and regulatory submissions. A general consensus was reached on the readiness of NGS to replace the in vivo adventitious virus detection assays and PCR assays, and to supplement or replace the in vitro cell-based assays, based on a suitable validation package.