Monkeypox (Mpox) has once again been designated a Public Health Emergency of International Concern (PHEIC) as of August, 2024. The severity of the disease is underscored by its significant mortality rate, and unfortunately, there are no targeted therapeutics currently available for this viral infection. Management relies on preventive measures and the use of existing smallpox vaccines due to their genetic similarity to the Mpox virus. Diagnosing a disease is a critical aspect of managing any health condition, and for a highly contagious viral infection like Mpox, it is essential to employ a specific and sensitive diagnostic approach. The lack of adequate diagnostic facilities in laboratories poses a significant challenge, hindering accurate diagnoses and the identification of underlying etiologies, particularly in low-resource settings. Current serology-based diagnostic tests lack specificity for the Mpox virus, leading to cross-reactivity with other orthopoxviruses. With the emergence of new viral variants, molecular and genomic diagnostic methods are far more reliable for accurately confirming Mpox infections. This review focuses on current diagnostic methods approved worldwide and the future challenges that need to be addressed to effectively control and mitigate the spread of Mpox.