Mycobacterium tuberculosis (MTB) is the pathogen that causes tuberculosis. It can invade all organs of the body, and pulmonary tuberculosis is the most common disease, which seriously endangers human health. This article reviews the mainstream detection techniques of MTB and their clinical applicability, focusing on analyzing the technical principles and performance differences of bacteriological examination, molecular biology detection, gene chips, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. It also discusses the advantages and disadvantages of different methods in terms of sensitivity, specificity, detection timeliness, and cost, and analyzes the causes of data heterogeneity in the literature. Finally, based on the limitations of various methods, a “stepwise combined detection” strategy is proposed, aiming to provide scientific basis for clinical selection of the optimal detection plan for different populations and resource conditions.