Analysis of bacterial DNA in synovial tissue of Tunisian patients with reactive and undifferentiated arthritis by broad-range PCR, cloning and sequencing

Abstract

Introduction Bacteria and/or their antigens have been implicated in the pathogenesis of reactive arthritis (ReA). Several studies have reported the presence of bacterial antigens and nucleic acids of bacteria other than those specified by diagnostic criteria for ReA in joint specimens from patients with ReA and various arthritides. The present study was conducted to detect any bacterial DNA and identify bacterial species that are present in the synovial tissue of Tunisian patients with reactive arthritis and undifferentiated arthritis (UA) using PCR, cloning and sequencing. Methods We examined synovial tissue samples from 28 patients: six patients with ReA and nine with UA, and a control group consisting of seven patients with rheumatoid arthritis and six with osteoarthritis (OA). Using broad-range bacterial PCR producing a 1,400-base-pair fragment from the 16S rRNA gene, at least 24 clones were sequenced for each synovial tissue sample. To identify the corresponding bacteria, DNA sequences were compared with sequences from the EMBL (European Molecular Biology Laboratory) database. Results Bacterial DNA was detected in 75% of the 28 synovial tissue samples. DNA from 68 various bacterial species were found in ReA and UA samples, whereas DNA from 12 bacteria were detected in control group samples. Most of the bacterial DNAs detected were from skin or intestinal bacteria. DNA from bacteria known to trigger ReA, such as Shigella flexneri and Shigella sonnei , were detected in ReA and UA samples of synovial tissue and not in control samples. DNA from various bacterial species detected in this study have not previously been found in synovial samples. Conclusion This study is the first to use broad-range PCR targeting the full 16S rRNA gene for detection of bacterial DNA in synovial tissue. We detected DNA from a wide spectrum of bacterial species, including those known to be involved in ReA and others not previously associated with ReA or related arthritis. The pathogenic significance of some of these intrasynovial bacterial DNAs remains unclear.

Clinical features and disease outcomes of undifferentiated arthritis in Thailand

“…Chlamydia surface antigen and RNA have been detected in the synovial tissue of reactive arthritis and/or Reiter’s syndrome; 22,23 however, recent data showed that it could be detected in the synovium of patients with undifferentiated oligoarthritis, RA and osteoarthritis 24 . Moreover, using the polymerase chain reaction (PCR) technique, bacterial DNA has been found in the synovial fluid and tissue in reactive arthritis and UA 25 . Albeit that infection could be related to UA, the major problem is that most patients (as in our study) cannot recall the history of infection preceding the onset of joint symptoms.…”

Detection and frequency ofChlamydia trachomatisDNA in synovial samples from Tunisian patients with reactive arthritis and undifferentiated oligoarthritis

“…Thirty-four patients were included in the study and had given their informed consent before arthrocentesis. Twentyeight of 34 samples analyzed in this study are identical to those from our previous study (Siala et al, 2008). All patients were attending one of three hospitals with rheumatological departments in Tunisia during the period of 2003-2007.…”

Prokaryotic diversity in continuous cropping and rotational cropping soybean soil

“…Cultivation-independent methodologies, particularly the sequence analysis of cloned 16S rRNA genes (16S rDNA), allowed extensive studies of the microbial diversity of ecosystems without the bias imposed by pureculture techniques (or at least with a different one), and have proven to be powerful tools for investigating the microbial diversity of environmental samples (Head et al, 1998). This method, used in many different environments, has identified new microorganisms that may be abundant or physiologically significant (Kennedy et al, 2008;Siala et al, 2008;Yoshida et al, 2008;Lakra et al, 2009).…”