Helicobacter pylori Mutations Detected by Next-Generation Sequencing in Formalin-Fixed, Paraffin-Embedded Gastric Biopsy Specimens Are Associated with Treatment Failure

摘要

Helicobacter pylori antibiotic resistance is widespread and increasing worldwide. Routine detection of H. pylori mutations that invoke antimicrobial resistance may be a useful approach to guide antimicrobial therapy and possibly avert treatment failure. In this study, formalin-fixed, paraffin-embedded (FFPE) gastric biopsy specimens from a cohort of individuals from northern Ohio in the United States were examined using a next-generation sequencing (NGS) assay to detect H. pylori mutations that are known to confer resistance to clarithromycin, levofloxacin, and tetracycline. From January 2016 to January 2017, 133 H. pylori-infected gastric biopsy specimens were identified histologically and subsequently analyzed by NGS to detect mutations in gyrA, 23S rRNA, and 16S rRNA genes. The method successfully detected H. pylori in 126 of 133 cases (95 sensitivity). Mutations conferring resistance were present in 92 cases (73), including 63 cases with one mutation (50) and 29 cases with mutations in multiple genes (23). Treatment outcomes were available in 58 cases. Sixteen of the 58 cases failed therapy (28). Therapy failure correlated with the number of mutated genes: no failure in cases with no mutations (0/15), 19 (5/27) failure in cases with one gene mutation, and 69 (11/16) failure in cases with more than one mutated gene. Common 235 rRNA mutations (A2142G or A2413G) were present in 88 (14/16) of failed cases as opposed to in only 10 (4/42) of eradicated cases (P < 0.001). This NGS assay can be used on remnant specimens collected during standard-of-care testing to detect mutations that correlate with increased risk of treatment failure. A prospective study is needed to determine if the risk of treatment failure can be decreased by using this assay to guide antibiotic therapy.