IJSR International Journal of Scientific Research 2277 - 8179 Indian Society for Health and Advanced Research ijsr-8-12-23486 Original Research Paper STUDY OF ASSOCIATION OF INTESTINAL PREVOTELLA IN EARLY RHEUMATOID ARTHRITIS Sabarish VS Dr. Dr Nandini K Dr. Dr Ashok P Dr. December 2019 8 12 01 02 ABSTRACT

Background: – Despite being highly prevalent and highly studied autoimmune disease, the etiology for the autoimmunity in Rheumatoid arthritis is not known. One of the recent advances being the role of gut microbiota. The primary objective of this study is to explore the association of Prevotella copri species in the gut microbiota in untreated Rheumatoid arthritis patients. AIMS AND OBJECTIVES:– To study the association of Prevotella copri in the gut microbiota in untreated Rheumatoid Arthritis patients MATERIAL AND METHODS: Stool samples were collected from controls and new onset, untreated Rheumatoid arthritis patients in reduced transport fluid (RTF) and were subjected to anaerobic culture in Kanamycin–Vancomycin blood agar to identify Prevotella species based on colony morphology and biochemical tests. Inoculated plates were incubated anaerobically by gas pak method for 72 hours. Also, broadband PCR was run on stool samples collected in RTF for detection of 16S RNA of Prevotella species and the samples which test positive were further subjected to specific PCR with another set of internal primers to detect 16S RNA of Prevotella copri. HiPurATM Stool DNA Purification Kit (#MB544 HIMEDIA) was used for extraction of bacterial genome. Separation of genomic DNA was done using agarose gel electrophoresis. 2 sets of primers were used among which one is universal primer for bacterial species i.e., 16S rDNA to validate the sample or DNA for bacterial genomic study. Another primer mix specific to Prevotella copri was procured from Helini Biosciences along with positive control for Prevotella copri to identify samples positive for Prevotella copri. Results:– Stool samples were collected from 30 cases and 25 healthy controls and were subjected to PCR and culture. Anaerobic culture showed no growth of Prevotella. PCR studies showed 19(63%) cases being positive for Prevotella copri nucleic acid whereas only 7(28%) samples were positive in controls. Conclusion: – Intestinal Prevotella copri was found in significant number of cases of Rheumatoid arthritis compared to controls (p value: 0.009) indicating the alteration in gut microbiota in cases, which could be the reason for priming of autoimmunity in Rheumatoid arthritis.