Detection of Chlamydophila pneumoniae in patients with arthritis: significance and diagnostic value

Abstract

The aim of this study was to assess the potential clinical implications of Chlamydophila pneumoniae in patients with acute and chronic arthritic diseases and to investigate whether blood monocytes might reflect a concomitant synovial or persistent systemic infection. C. pneumoniae was investigated with advanced PCR and reverse transcriptase (RT) PCR techniques targeting different genes and combined with cell line cultures, in synovial fluid (SF) and peripheral blood mononuclear cell (PBMC) specimens collected from 28 patients with arthritis. Five out of twenty-eight patients (17.8%) were found to have C. pneumoniae DNA in either SF or PBMC specimens. Their diagnosis was reactive arthritis (ReA), S.A.P.H.O syndrome, psoriatic arthritis, undifferentiated oligoarthritis (UOA) and ankylosing spondylitis (AS). Specimens from patients with UOA and AS had also mRNA transcripts but those from AS yielded C. pneumoniae growth after co-culture. Moreover, C. pneumoniae DNA levels measured by Real-Time PCR (LightCycler) were higher in PBMC specimens compared to those found in SF at the end of antibiotic treatment. C. pneumoniae may have a role as triggering factor also in chronic arthritides including AS. The combined use of culture and molecular tools increases detection rates and improves the overall sensitivity, suggesting their potential use to detect C. pneumoniae. The different kinetics of bacterial DNA at both peripheral and synovial levels should be taken into consideration when monitoring and evaluating the effectiveness of antibiotic treatment.

Detection of Chlamydophila pneumoniae and human herpesvirus 8 in primary cutaneous anaplastic large-cell lymphoma: a case report

“…DNA and RNA were extracted as previously described [10,11], and assayed by PCR. A fraction of PBMCs was instead centrifuged, suspended in RPMI 1640 medium (Gibco, Invitrogen, Carlsbad, CA) and co-cultured up to 144 hours with Hep-2 cell line (ATCC CCL-23), to increase the number of bacterial inclusions [11]. Cultured samples were then processed as done for fresh uncultured cells.…”

“…Cultured samples were then processed as done for fresh uncultured cells. Analyses included nested-PCR and PCR after retrotranscription (RT-PCR), using primer sets targeting 16S rRNA, outer membrane protein (ompA/MOMP), and HsP-60 genomic regions of C. psittaci , C. pneumoniae , and C. trachomatis [11]. Amplification fragments, corresponding to C. pneumoniae 16S rRNA gene, were found both in tumor tissue and in co-cultured PBMCs (Figure  3).…”

HPV genotypes distribution in Chlamydia trachomatis co-infection in a large cohort of women from north-east Italy

“…The sensitivity of each run was defined as the lowest dilution of DNA (2|10 24 ng/ml, corresponding to 1 genome copies/ml) detected by the 7900HT instrument (Applied Biosystems) (Contini et al, 2011). Standard curve equations were used to calculate the absolute number of gene mRNA copies.…”

The Role of Chlamydia and Chlamydophila Infections in Reactive Arthritis

“…The persistence of chlamydial infection (38,39) may amplify joint inflammation and delay the clearance of the pathogen (34). The atypical reticulate bodies were postulated to explain why this infection escapes detection by conventional methods (40) and escapes the immune surveillance (41).…”

“…Previous RT-PCR-based studies demonstrated that DNA from these organisms is often present in synovial tissue in ReA patients (33), but the simple presence of chlamydial DNA by PCR does not define a clinical syndrome or disease course and it does not provide insight into bacterial viability or metabolic activity (34). Therefore, the association of a culture and the molecular evidence that these organisms or their molecular components are found in the synovia of affected individuals is important (30).…”