The article reviews the evolution of laboratory diagnosis of tuberculosis (TB), emphasizing the importance of accurate and timely diagnosis in combating the disease. TB, caused by the Mycobacterium tuberculosis complex, remains a global public health concern, exacerbated by the emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains. The review highlights the limitations of traditional diagnostic methods, such as microscopic examination and culture, and the advantages of molecular assays like LPA and Xpert MTB/RIF in rapid and accurate TB diagnosis, including drug resistance detection. The integration of multiple diagnostic methods is crucial for effective TB management. The review also discusses the challenges posed by the COVID-19 pandemic, which disrupted TB diagnosis and treatment access, leading to a decline in TB cases diagnosed in 2020. The article emphasizes the need for improved diagnostic technologies, increased accessibility, and the adoption of advanced methods to enhance TB diagnosis accuracy and efficiency. It also addresses the limitations of traditional methods, such as the Ziehl-Neelsen (ZN) staining, and the potential of newer techniques like fluorescent staining, LED fluorescence microscopy, and automated methods in improving TB diagnosis. The review highlights the importance of molecular tests in identifying drug resistance mutations and their role in guiding appropriate treatment. The study also discusses the effectiveness of various decontamination methods, such as NaOH-NALC and chlorhexidine, in improving sample preparation for TB diagnosis. The article concludes that a combination of traditional and molecular methods is essential for accurate TB diagnosis and effective control of the disease.The article reviews the evolution of laboratory diagnosis of tuberculosis (TB), emphasizing the importance of accurate and timely diagnosis in combating the disease. TB, caused by the Mycobacterium tuberculosis complex, remains a global public health concern, exacerbated by the emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains. The review highlights the limitations of traditional diagnostic methods, such as microscopic examination and culture, and the advantages of molecular assays like LPA and Xpert MTB/RIF in rapid and accurate TB diagnosis, including drug resistance detection. The integration of multiple diagnostic methods is crucial for effective TB management. The review also discusses the challenges posed by the COVID-19 pandemic, which disrupted TB diagnosis and treatment access, leading to a decline in TB cases diagnosed in 2020. The article emphasizes the need for improved diagnostic technologies, increased accessibility, and the adoption of advanced methods to enhance TB diagnosis accuracy and efficiency. It also addresses the limitations of traditional methods, such as the Ziehl-Neelsen (ZN) staining, and the potential of newer techniques like fluorescent staining, LED fluorescence microscopy, and automated methods in improving TB diagnosis. The review highlights the importance of molecular tests in identifying drug resistance mutations and their role in guiding appropriate treatment. The study also discusses the effectiveness of various decontamination methods, such as NaOH-NALC and chlorhexidine, in improving sample preparation for TB diagnosis. The article concludes that a combination of traditional and molecular methods is essential for accurate TB diagnosis and effective control of the disease.