AIM To investigate the intestinal luminal microbiota (LM) and mucosa-associated microbiota (MAM) in Chinese patients with functional gastrointestinal disorders (FGIDs) and examine the association between these communities and the expression of toll-like receptor (TLR) 2 and TLR4. METHODS Thirty-two Chinese subjects who suffered from symptoms of FGIDs, as confirmed by gastroenterologists, were enrolled in this study. Fresh faecal samples and descending colonic mucosal biopsies were collected from the subjects before (faecal) and during (mucosal) ﬂexible colonoscopy. For analysis of the samples, we performed high-throughput sequencing of the V3-V4 region of the 16S rRNA gene and reverse transcription (RT)-PCR to detect the expression of colonic TLR2 and TLR4. Differences in the stool and mucosal microbiota were examined and a correlation network analysis was performed. RESULTS The microbiota of faecal samples was significantly more diverse and richer than that of the mucosal samples, and the LM and MAM populations differed signiﬁcantly. TLR2 expression showed a significant positive correlation with TLR4 expression. In the MAM samples, the genera Faecalibacterium and Ruminococcus, which belong to the family Ruminococcaceae, were inversely correlated with TLR4 expression (r = -0.45817, P = 0.0083 and r = -0.5306, P = 0.0018, respectively). Granulicatella, which belongs to Carnobacteriaceae, and Streptococcus, which belongs to Streptococcaceae, were inversely correlated with TLR2 expression (r = -0.5573, P = 0.0010 and r = -0.5435, P = 0.0013, respectively). In the LM samples, examination at phylum, class, or order level revealed no correlation with TLR4 expression. Faecalibacterium, which belongs to Ruminococcaceae, and Streptococcus, which belongs to Streptococcaceae, were inversely correlated with TLR2 expression (r = -0.5743, P = 0.0058 and r = -0.3905, P = 0.0271, respectively). CONCLUSION Microbial compositions of LM and MAM in Chinese patients with FGIDs are different. Expression of TLRs may be affected by the type of bacteria that are present in the gut.