Abstract
Chronic suppurative otitis media (CSOM) is a prevalent condition with global health implications due to its impact on hearing and quality of life. Conventional treatments often fail because of bacterial biofilms and antimicrobial resistance. Effective treatment of CSOM depends on the precise determination of the middle ear microbiota; however, current microbial detection methods do not meet this need. Postoperative reperforation may compromise surgical outcomes. If the risk of perforation can be predicted immediately after surgery, sensitive antibiotics could be administered proactively for early intervention to optimize treatment efficacy. This study introduces 2b-RAD sequencing for the microbiome (2b-RAD-M), a novel technology designed to provide a comprehensive profile of the CSOM microbiota and identify diagnostic biomarkers that predict postoperative outcomes. We analyzed ear swabs from patients with postoperative perforation (PO), nonperforation (NPO), and otosclerosis (CON) using microbial diversity, relative abundance, and composition analyses. Bacillus_A_bombysepticus and Pseudomonas aeruginosa were identified as potential biomarkers, with Bacillus_A_bombysepticus demonstrating superior diagnostic accuracy (area under curve (AUC) = 0.92) compared to P. aeruginosa (AUC = 0.25). Functional predictions revealed that biological activities related to gene regulation, substance metabolism, and DNA repair were more prominent in the PO group. This study offers new insights into CSOM pathogenesis and progression, proposing Bacillus_A_bombysepticus as a novel biomarker for predicting postoperative outcomes that can indicate an increased risk of tympanic membrane reperforation for the first time. KEY POINTS: 2b-RAD-M technology enables comprehensive CSOM microbiota profiling and biomarker identification. Bacillus_A_bombysepticus (AUC = 0.92) outperforms Pseudomonas aeruginosa in diagnostic accuracy. Bacillus_A_bombysepticus predicts postoperative tympanic membrane reperforation via functional activity analysis.