Nested Culture Method Improves Detection of Fusobacterium from Stool in Patients with Ulcerative Colitis

Abstract

Fusobacterium varium is an elusive pathogenic factor in ulcerative colitis (UC); conventional methods of fecal culture rarely recover F. varium. We have developed a nested culture method to recover Fusobacterium and we used it to investigate whether F. varium could be isolated from UC patients. We enrolled 50 consecutive patients in this study; 26 received combination antibiotic therapy that included amoxicillin, tetracycline, and metronidazole (ATM) for 2 weeks and were thus assigned to the ATM group, and the remaining 24 were assigned to the non-ATM group and did not receive any antibiotics. Stool samples were added to 10 ml of GAM broth that contained neomycin and crystal violet. The samples were vortexed and incubated under anaerobic conditions. The preincubated broth was streaked onto a Fusobacterium-selective agar plate and then incubated under anaerobic conditions. The species of the colonies isolated were identified using the Vitek Automated system and PCR analysis. We recoverd F. varium from 7 of the 24 non-ATM patients (29.2%) and none from the ATM patients (0%) (P = 0.0035). All of the F. varium isolates were susceptible to ATM. This study suggests that the recovery of F. varium is related to UC, which aligns with results from previous studies that used mucosal culture, immunostaining, real-time PCR, and serological studies.