| Objective: To characterize the gut microbiota in patients with Graves’ disease andassess the diversity and structure of intestinal flora in Graves disease patients comparedwith healthy individuals.Methods: Fresh fecal samples from14patients with Graves’ disease and13healthy individuals were collected. The samples were collected and analyzed in3serials.Fecal bacterial genome DNA was extracted with a DNA-extraction kit. After PCRamplication with universal primers targeting V3region of the16S rRNA gene,denaturing gradient gel electrophoresis (PCR-DGGE) was employed to characterize theoverall intestinal microbiota composition. Distinct bands were excised and cloned forsequencing. The sequence was compared directly with that in GenBank by Blast searchin NCBI.Result: The dominant gut microbiota of patients with Graves’ disease was founddifferent from healthy groups. DNA fingerprints was analyzed with Quantity One4.6.2software. In the first experimental group, there was no significant difference in thediversity of intestinal flora in patients with Graves’ disease from that in health subjects,the Dice similarity coefficient within subjects was greater than that between subjects.The dendrogram was constructed based on analysis of similarity score and cluster fromDGGE profiles by UPGMA method. Statistically significant clustering profiles wereconstructed in patients with Graves’ disease and in healthy subjects except sample G8and G10; In the second experimental serial, the diversity of intestinal flora in patientswith Graves’ disease was not different from health control subjects, Dice similaritycoefficient within subjects was greater than the similarity between subjects. But thedendrogram showed that the Graves’ disease patients and healthy groups formedstatistically significant clustering profiles except sample G6. In the third experimentalgroup, the diversity of intestinal flora in patients with Graves’ disease was significantly different from health controls (P＜0.05), the Dice similarity coefficient within subjectswas greater than between subjects. The dendrogram analysis found the Graves’ diseasepatients and heathy groups formed statistically significant clustering profiles, exceptsample C11. By choosing one dominant bands in patients with Graves’ diseasecompared healthy individuals, this band was sequenced and identified as Prevotellaspecies.Conclusion: No significant difference in the diversity of intestinal microbiotabetween patients with Graves disease and healthy individuals except the abundance ofsome microbiota in patients with Graves disease. Increases in Prevotella species mightbe closely related to pathogenesis of Graves disease, and need further investigation. |
