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Yuxin Fan,1– 3 Jingjing Chen,1,4 Meixiao Liu,1,2 Xin Xu,1,2 Yu Zhang,1– 3 Peng Yue,1,4 Wenjing Cao,1,4 Zhenhua Ji,1,2 Xuan Su,1,2 Shiyuan Wen,1– 3 Jing Kong,1,2 Guozhong Zhou,1,2 Bingxue Li,1,2 Yan Dong,1,2 Aihua Liu,1,3,4 Fukai Bao1– 3 1The Institute for Tropical Medicine, Faculty of Basic Medical Sciences, Kunming Medical University, Kunming, 650500, People’s Republic of China; 2Department of Microbiology and Immunology, Kunming Medical University, Kunming, 650030, People’s Republic of China; 3Yunnan Province Key Laboratory of Children’s Major Diseases Research, The Affiliated Children Hospital, Kunming Medical University, Kunming, 650030, People’s Republic of China; 4Department of Biochemistry and Molecular Biology, Kunming Medical University, Kunming, 650030, People’s Republic of ChinaCorrespondence: Aihua Liu; Fukai Bao, The Institute for Tropical Medicine, Faculty of Basic Medical Sciences, Kunming Medical University, Kunming, 650500, People’s Republic of China, Email liuaihua@kmmu.edu.cn; baofukai@kmmu.edu.cnAbstract: Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (MTB) infection, which has seriously endangered human health for many years. With the emergence of multidrug-resistant and extensively drug-resistant MTB, the prevention and treatment of TB has become a pressing need. Early diagnosis, drug resistance monitoring, and control of disease transmission are critical aspects in the prevention and treatment of TB. However, the currently available diagnostic technologies and drug sensitivity tests are time consuming, and thus, it is difficult to achieve the goal of early diagnosis and detection drug sensitivity, which results in limited control of disease transmission. The development of molecular testing technology has gradually achieved the vision of rapid and accurate diagnosis of TB. Droplet digital PCR (ddPCR) is an excellent nucleic acid quantification method with high sensitivity and no need