Analysis on the pharyngeal microbiota in patients with laryngopharyngeal reflux disease()

OBJECTIVE(S): In this study, the laryngopharynx microbiome alterations were characterized after proton pump inhibitor treatment in patients with Laryngopharyngeal Reflux Disease (LPRD) and healthy people. The potential outcome-predictive biomarker was explored. METHODS: Patients with LPRD and healthy controls were enrolled. The composition of their laryngopharynx microbiota was analyzed both by traditional plate count of the main bacterial groups and PCR amplification followed by denaturing gradient gel electrophoresis. Shannon-Wiener index and evenness index based on Dice index were used to assess the bacterial diversity. Droplet digital PCR was used to determine the total bacterial RNA and relative abundance of Klebsiella oxytoca. Receiver operating characteristic curve was plotted to explore the potential of Klebsiella oxytoca as an outcome-predictive biomarker. RESULTS: A total of 29 LPRD cases and 28 healthy subjects were enrolled. The composition of the laryngopharynx microbiota was almost similar, except Klebsiella oxytoca. The cluster analysis showed that the similarity between healthy and treatment-effective groups, as well as pretreatment and treatment-invalid groups, was close. Statistical analysis showed that there were differences in the diversity index and richness among the healthy, treatment-effective, pretreatment and treatment-invalid groups. The abundance of Klebsiella oxytoca in the treatment-effective LPRD group was lower than that of the treatment-invalid LPRD group. The abundance of Klebsiella oxytoca can distinguish treatment-effective and -invalid groups (AUC = 0.859) with a sensitivity of 77.78% and specificity of 90.91%. CONCLUSION: There were differences in the diversity of cecal contents microbial community between treatment-invalid and treatment-effective LPRD groups. Klebsiella oxytoca has potential to distinguish treatment outcomes. LEVEL OF EVIDENCE: How common is the problem? Level 1. Is this diagnostic or monitoring test accurate? (Diagnosis) Level 4. What will happen if we do not add a therapy? (Prognosis) Level 5. Does this intervention help? (Treatment Benefits) Level 4. What are the COMMON harms? (Treatment Harms) Level 4. What are the RARE harms? (Treatment Harms) Level 4. Is this (early detection) test worthwhile?(Screening) Level 4.