Development of a Four Autophagy-related Gene Signature for Active Tuberculosis Diagnosis

Baoyan Ren¹Feng Jia²Qixun Fang¹Jingping Xu^1,3Kangfeng Lin^1,3Renhui Huang¹Zhenqiong Liu²Xingan Xing^1,3*

• ¹Yaneng BIOscience (Shenzhen) Co., Ltd, Guangdong, China
• ²Jiangxi Chest Hospital, Nanchang, Jiangxi Province, China
• ³South China University of Technology, Guangzhou, China

Tuberculosis (TB) diagnostics urgently require non-sputum biomarkers to address the limitations of conventional methods in distinguishing active TB (ATB) from latent infection (LTBI) and confounding diseases. By leveraging autophagy-a key host defense pathway against Mycobacterium tuberculosis (Mtb), we identified a novel four-gene autophagy-related signature (CASP1, FAS, TRIM5, C5) through transcriptomic profiling of peripheral blood and machine learning-driven selection in selection dataset. This signature demonstrated robust diagnostic accuracy across multiple evaluation datasets: Area Under the Curve (AUC) 0.83-0.98 for ATB vs. LTBI and 0.82-0.94 for ATB vs. healthy controls (HCs). Crucially, it maintained high specificity (AUC 0.89-0.90) against TB-mimicking diseases (ODs). RT-qPCR validation in whole blood samples confirmed elevated expression in ATB, while a support vector machine (SVM) model achieved promising differentiation (AUC 0.86 for ATB vs. LTBI and AUC 0.99 for ATB vs. HCs). Overall, our findings yielded a four-gene signature in whole blood that is robustly diagnostic for ATB, validated across multiple evaluation datasets and clinical samples. The autophagy-driven specificity and PCR-compatible design of this signature offer a blood-based, costeffective strategy to enhance TB detection, addressing WHO-aligned diagnostic needs.