Abstract: Objective To evaluate the application of TB laboratory detection technology in Liaoning Province from 2016 to 2022, and to provide scientific basis for further improving the detection rate of Mycobacterium tuberculosis in the province. Methods The medical records of registered tuberculosis patients in Liaoning Province from 2016 to 2022 were collected from the "Tuberculosis Information Management System" in the "China Disease Prevention and Control Information System" subsystem. Statistical analysis was performed for sputum coating, sputum culture, and molecular biology testing. Results From 2016 to 2022, a total of 152 778 patients with pulmonary tuberculosis were registered in Liaoning Province. The detection rate of sputum smear microscopy was 98.03% (149 775/152 778), the detection rate of sputum culture was 20.72% (31 661/152 778), and the detection rate of molecular biology testing was 20.21% (30 737/152 778). From 2018 to 2022, the rate of molecular biological detection showed an increasing trend (χ2trend=7 104.466, P<0.01), while from 2016 to 2021, the detection rate of sputum culture showed an increasing trend, with statistical significance (χ2trend=3,068.701, P<0.01). The sputum smear detection rate showed a downward trend（χ2trend=689.913, P<0.01). . There were significant differences in the results of sputum smear microscopy, sputum culture, and molecular biology testing, as confirmed by the McNemar test (P<0.01). The positive rate of pathogenic academics increased from 26.27% in 2016 to 51.55% in 2022, showing a yearly upward trend (χ2trend=5 262.863, P<0.01), with significant differences between each year (χ2=5 686.935, P<0.01). Among pulmonary tuberculosis patients with positive pathogenic microorganisms, the proportion of sputum smear-positive cases decreased from 94.32% to 52.36%, showing a downward trend (χ2trend=5 010.104, P<0.01). The proportion of culture-positive cases increased from 5.68% in 2016 to 12.83% in 2022, showing an upward trend (χ2trend=122.501, P<0.01). In Liaoning Province, molecular biology testing has been carried out since 2018, and the proportion of molecular biology-positive cases increased from 11.51% to 34.81%, showing an increasing trend (χ2trend=1 969.326, P<0.01). The number of positive patients in molecular biological tests in municipal hospitals accounted for 18.69% (8 386/44 778) of etiological positive patients, while the number of positive patients in county-level hospitals accounted for 13.61% (2 439/17 924) of etiological positive patients, with significant differences (χ2=231.594, P<0.01). Conclusions The implementation of molecular biology testing for tuberculosis in Liaoning Province is one of the main measures to improve the positive rate of etiology, and it helps to diagnose tuberculosis patients timely and accurately