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Broad-range rDNA PCR provides an alternative, cultivation-independent approach for identifying bacterial DNA in reactive and other form of arthritis. The aim of this study was to use broad-range rDNA PCR targeting the 16S rRNA gene in patients with reactive and other forms of arthritis and to screen for the presence of DNA from any given bacterial species in synovial fluid (SF) samples.    Methods  We examined the SF samples from a total of 27 patients consisting of patients with reactive arthritis (ReA) (n = 5), undifferentiated arthritis (UA) (n = 9), rheumatoid arthritis (n = 7), and osteoarthritis (n = 6) of which the latter two were used as controls. Using broad-range bacterial PCR amplifying a 1400 bp fragment from the 16S rRNA gene, we identified and sequenced at least 24 clones from each SF sample. To identify the corresponding bacteria, DNA sequences were compared to the EMBL (European Molecular Biology Laboratory) database.    Results  Bacterial DNA was identified in 20 of the 27 SF samples (74, 10%). Analysis of a large number of sequences revealed the presence of DNA from more than one single bacterial species in the SF of all patients studied. The nearly complete sequences of the 1400 bp were obtained for most of the detected species. DNA of bacterial species including  Shigella  species,  Escherichia  species, and other coli-form bacteria as well as opportunistic pathogens such as  Stenotrophomonas maltophilia  and  Achromobacter xylosoxidans  were shared in all arthritis patients. Among pathogens described to trigger ReA, DNA from  Shigella sonnei  was found in ReA and UA patients. We also detected DNA from rarely occurring human pathogens such as  Aranicola  species and  Pantoea ananatis . We also found DNA from bacteria so far not described in human infections such as  Bacillus niacini ,  Paenibacillus humicus ,  Diaphorobacter  species and uncultured bacterium genera incertae sedis OP10.    Conclusions  Broad-range PCR followed by cloning and sequencing the entire 16S rDNA, allowed the identification of the bacterial DNA environment in the SF samples of arthritic patients. We found a wide spectrum of bacteria including those known to be involved in ReA and others not previously associated with arthritis.

arthritis research and therapy

Broad-range PCR, cloning and sequencing of the full 16S rRNA gene for detection of bacterial DNA in synovial fluid samples of Tunisian patients with reactive and undifferentiated arthritis