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Detection of bacterial DNA in Latin American patients with reactive arthritis by polymerase chain reaction and sequencing analysis
Authors:Cuchacovich Raquel  Japa Shankar  Huang Wen Qun  Calvo Armando  Vega Luis  Vargas Ruben Burgos  Singh Ranju  Flores Diana  Castro Ivette  Espinoza Luis R
Institution:Department of Medicine, LSU Health Science, Center, New Orleans, Louisiana 70112, USA.
Abstract:OBJECTIVE: Bacteria and/or their antigens are thought to play a role in the pathogenesis of reactive arthritis (ReA). Polymerase chain reaction (PCR) using the 16S ribosomal RNA-PCR method was used to identify bacterial DNA in synovial fluid (SF) and tissue (ST) in a well defined group of patients with chronic ReA. In addition, species found were identified by means of sequence analysis. METHODS: We examined 15 ST and 5 SF samples of 15 patients with ReA, 5 ST samples of 5 patients with osteoarthritis (OA), and 8 SF from 8 patients with closed traumatic knee injuries using a nested PCR with universal 16S rRNA primers. In addition, a nested PCR was developed to detect DNA sequences of Salmonella sp. and Mycoplasma sp. Automated sequencing and comparative data analysis (GenBank) were also performed to identify the species. RESULTS: Bacterial DNA was identified in 8 cases, 5 ST and 3 SF; Chlamydia trachomatis (n = 2), Pseudomonas sp. (n = 3), and Bacillus cereus (n = 2) were the most common microorganisms identified. A variety of microorganisms including Clostridium sp., Lactobacillus sp., Pseudomonas migulae, P. fluorescens, and P. putida, and Neisseria meningitidis serogroup B were also identified. In half of the cases (4/8) 2 to 3 bacterial antigens were identified simultaneously. CONCLUSION: Bacterial DNA is present in the joints in patients with chronic ReA. A wide spectrum of bacteria including some not previously associated with ReA were identified. Further studies are needed to establish their exact role in the pathogenesis of ReA and related arthritides.
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