This review provides an in-depth examination of the epidemiology, pathogenesis, and diagnosis of Monkeypox (Mpx), a zoonotic disease caused by the Orthopoxvirus genus. The authors highlight the increasing risk of Mpx transmission to humans due to urbanization and increased interaction between humans and wild animals. They discuss the molecular mechanisms of viral replication, including the initiation of DNA synthesis and the production of viral proteins, as well as the host immune response, which involves both innate and adaptive mechanisms. The review also covers diagnostic methods such as nucleic acid amplification testing (NAAT), electron microscopy, virus isolation and culture, serology, and whole-genome sequencing (WGS). The importance of public health implications, including vaccination, surveillance, and emergency preparedness, is emphasized to prevent and manage potential outbreaks. The 2022-2023 Mpx outbreak, which spread rapidly across Europe, is noted for its unique transmission patterns and demographic shift, particularly among men who have sex with men. The review underscores the need for continued research, surveillance, and adaptive public health interventions to address the evolving threat of Mpx.This review provides an in-depth examination of the epidemiology, pathogenesis, and diagnosis of Monkeypox (Mpx), a zoonotic disease caused by the Orthopoxvirus genus. The authors highlight the increasing risk of Mpx transmission to humans due to urbanization and increased interaction between humans and wild animals. They discuss the molecular mechanisms of viral replication, including the initiation of DNA synthesis and the production of viral proteins, as well as the host immune response, which involves both innate and adaptive mechanisms. The review also covers diagnostic methods such as nucleic acid amplification testing (NAAT), electron microscopy, virus isolation and culture, serology, and whole-genome sequencing (WGS). The importance of public health implications, including vaccination, surveillance, and emergency preparedness, is emphasized to prevent and manage potential outbreaks. The 2022-2023 Mpx outbreak, which spread rapidly across Europe, is noted for its unique transmission patterns and demographic shift, particularly among men who have sex with men. The review underscores the need for continued research, surveillance, and adaptive public health interventions to address the evolving threat of Mpx.