This review provides an in-depth analysis of the pathogenesis, diagnosis, and public health implications of Mpox, caused by the Orthopoxvirus genus. Mpox is a zoonotic disease primarily affecting humans and animals, with transmission mainly from animals to humans, especially in endemic regions of Central and West Africa. Human-to-human transmission is also possible, particularly through close contact. The virus is transmitted through direct contact with infected animals, bodily fluids, or contaminated objects. Mpox has become an emerging public health threat due to increased urbanization and human-wildlife interactions. The review discusses the molecular mechanisms of viral replication, including the role of viral DNA polymerase and the production of viral proteins essential for replication. It also explores the host immune response, highlighting the roles of innate and adaptive immunity in combating Mpox. The virus has evolved strategies to evade immune detection, such as modulating host signaling pathways and interfering with cytokine signaling. Diagnostic methods for Mpox include nucleic acid amplification testing (NAAT), virus isolation and culture, electron microscopy, serology, and whole-genome sequencing (WGS). Each method has its strengths and limitations, with WGS being the most precise for differentiating Mpox from other orthopoxviruses. However, the availability and accessibility of these diagnostic tools vary, and challenges remain in ensuring timely and accurate diagnosis. Public health implications of Mpox include the need for vaccination, surveillance, and preparedness to prevent and manage outbreaks. Vaccination remains a key strategy, with live-attenuated vaccines derived from vaccinia virus showing effectiveness. Surveillance systems are crucial for early detection and outbreak response, while international collaboration is essential for managing the global spread of the virus. The review emphasizes the importance of continued research, surveillance, and adaptive public health interventions to address the evolving threat of Mpox.This review provides an in-depth analysis of the pathogenesis, diagnosis, and public health implications of Mpox, caused by the Orthopoxvirus genus. Mpox is a zoonotic disease primarily affecting humans and animals, with transmission mainly from animals to humans, especially in endemic regions of Central and West Africa. Human-to-human transmission is also possible, particularly through close contact. The virus is transmitted through direct contact with infected animals, bodily fluids, or contaminated objects. Mpox has become an emerging public health threat due to increased urbanization and human-wildlife interactions. The review discusses the molecular mechanisms of viral replication, including the role of viral DNA polymerase and the production of viral proteins essential for replication. It also explores the host immune response, highlighting the roles of innate and adaptive immunity in combating Mpox. The virus has evolved strategies to evade immune detection, such as modulating host signaling pathways and interfering with cytokine signaling. Diagnostic methods for Mpox include nucleic acid amplification testing (NAAT), virus isolation and culture, electron microscopy, serology, and whole-genome sequencing (WGS). Each method has its strengths and limitations, with WGS being the most precise for differentiating Mpox from other orthopoxviruses. However, the availability and accessibility of these diagnostic tools vary, and challenges remain in ensuring timely and accurate diagnosis. Public health implications of Mpox include the need for vaccination, surveillance, and preparedness to prevent and manage outbreaks. Vaccination remains a key strategy, with live-attenuated vaccines derived from vaccinia virus showing effectiveness. Surveillance systems are crucial for early detection and outbreak response, while international collaboration is essential for managing the global spread of the virus. The review emphasizes the importance of continued research, surveillance, and adaptive public health interventions to address the evolving threat of Mpox.