Comparison of synovial tissue and synovial fluid as the source of nucleic acids for detection of Chlamydia trachomatis by polymerase chain reaction

Abstract

Objective. Difficulties in detecting Chlamydia trachomatis in human joints by polymerase chain reaction (PCR) may be related to whether synovial tissue or synovial fluid (SF) is used as the source of DNA in PCR amplification. In this study, a new PCR assay was developed and used to compare chlamydial DNA in paired samples of SF and synovial tissue from patients with arthritis.Methods. The PCR assay targeted the ribosomal RNA operons, which are present in 2 copies on the C trachomatis chromosome. DNA from several relevant bacteria and chlamydial serovars was used for testing this screening system. The detection of chlamydial DNA in nucleic acid preparations from matched samples of SF and synovial tissue was compared by PCR assay. Samples were obtained from 55 patients, including patients with reactive arthritis, Reiter's syndrome, and other arthropathies.Results. Testing of the PCR screening system confirmed it to be highly specific and sensitive. Use of this assay to screen DNA from SF and synovial tissue samples showed that 29 (53%) of 55 synovial tissue preparations were positive for chlamydial DNA, but only 16 (29%) of the matched SF samples from these 29 patients were similarly positive. Five (9%) of 55 SF samples, but not their tissue counterparts, were positive for chlamydial DNA by PCR.Conclusion. Detection of chlamydial DNA in the joints of patients by PCR gives positive results more often when synovial tissue rather than SF is the source of target nucleic acids. Although synovial tissue is the source of choice for the most reliable determination of chlamydia in the joint, both synovial tissue and SF should be assayed if possible.

Diagnosis of Chlamydia trachomatis in patients with reactive arthritis and undifferentiated spondyloarthropathy

“…A study published in 2009 reported that C. trachomatis DNA in ReA and uSpA patients is rather high (100% and 64%, respectively) [13], while other studies reported relatively low frequencies (25%) [11]. Thus, published data on the use of PCR for the detection of intraarticular C. trachomatis varies greatly in patients with ReA and uSpA [10,33]. Since no results are available on inter-laboratory variability, these differences cannot be simply attributed to varying prevalence, as they may reflect different sensitivities and specificities of the employed PCR.…”

Early cytokine profiles in the joint define pathogen clearance and severity of arthritis in Chlamydia‐induced arthritis in rats

“…Besides the gene sequence for 16S rRNA, Bas et al (26) detected a major outer membrane protein (MOMP) gene sequence and 2 chlamydial plasmid sequences. Several investigators then reported obtaining positive PCR results for 16S rRNA (27,28). Gerard et al (29) took synovial specimens that tested positive for 16S rRNA by PCR and then further identified chlamydial gene products (glyQS, r-protein S5 and L5, and Hsp60) within a majority of these biopsy specimens.…”

Diagnosing reactive arthritis: Role of clinical setting in the value of serologic and microbiologic assays

“…In such a case, it has been shown that Chlamydia trachomatis is the triggering pathogen for the arthritis in ϳ50% of patients (18,21,22), resulting in a pretest probability for Chlamydia-induced arthritis of 50%. The pretest probability for Chlamydia-induced arthritis in patients presenting with a clinical picture compatible with ReA but without a symptomatic preceding infection has been estimated at ϳ12% (18) and 22% (3) if the calculation is based on serologic findings or detection of Chlamydia in the urogenital tract, and at ϳ30% (23)(24)(25) if based on polymerase chain reaction (PCR) methods to look for Chlamydia trachomatis in synovial fluid. Therefore, calculations are made for the different pretest probabilities of 12% and 30%.…”

“…Only small amounts of synovial fluid or membrane are necessary for such a test. In patients with undifferentiated arthritis but with a joint pattern compatible with ReA, chlamydial DNA has been detected by PCR in ϳ30% of cases in different countries (23)(24)(25). However, chlamydial DNA was also found in control groups such as patients with rheumatoid arthritis (25) or healthy subjects (30).…”