Tuberculosis (TB) remains one of the leading causes of death from a single infectious agent worldwide, a burden further exacerbated by HIV co-infection and the increasing prevalence of drug-resistant strains. Although a wide range of laboratory diagnostic methods are currently available, their applicability, implementation, and clinical impact vary substantially across healthcare settings with different levels of complexity and resources. This review provides a comprehensive overview of the main laboratory diagnostic methods for active and latent TB, emphasizing their clinical applicability, implementation challenges, and role within integrated diagnostic strategies. Conventional approaches, such as smear microscopy and culture, are discussed alongside modern diagnostic technologies, including automated nucleic acid amplification tests (NAATs), loop-mediated isothermal amplification (LAMP), line probe assays (LPAs), next-generation sequencing (NGS), and lateral flow assays, highlighting their strengths and limitations in distinct epidemiological and operational contexts. Unlike existing WHO guidelines and prior reviews that predominantly focus on test performance and recommendation status, this review adopts an implementation-oriented perspective, critically examining diagnostic methods in light of real-world constraints, regional disparities, and evidence gaps. Particular attention is given to limitations related to laboratory infrastructure, biosafety, workforce capacity, and sustainability, as well as to under-addressed areas such as latent TB, metagenomic approaches, and the investigation of co-pathogens. By integrating WHO guidance with contextual and operational considerations, this review aims to support rational test selection and the development of flexible, integrated diagnostic workflows tailored to local health system capacity, patient populations, and clinical scenarios, thereby strengthening the effectiveness and equity of TB diagnostic strategies.

Tuberculosis (TB) remains one of the leading causes of death from a single infectious agent worldwide, particularly aggravated by HIV co-infection and the increasing burden of drug-resistant strains. This review provides a comprehensive overview of current la-boratory diagnostic methods for active and latent TB, emphasizing their clinical applica-bility across different healthcare settings, diagnostic performance, and implementation in integrated testing workflows. Conventional methods, such as smear microscopy and cul-ture, are discussed alongside modern diagnostic approaches, including automated nucle-ic acid amplification tests (NAATs), loop-mediated isothermal amplification (LAMP), line probe assays (LPA), next-generation sequencing (NGS), and lateral flow assays for the di-agnosis of TB in specific clinical contexts. The strengths and limitations of each method are critically evaluated according to infrastructure level, resource availability, and epide-miological scenario. While traditional techniques remain useful in selected settings, mo-lecular technologies provide higher sensitivity, shorter turnaround times, and expanded capacity for drug resistance detection. The integration of complementary diagnostic strat-egies into hybrid testing algorithms is essential to optimize resource use, ensure diagnos-tic accuracy, promote equitable access, and enable early treatment initiation, thereby sup-porting effective TB control.