Optimized testing for C. trachomatis DNA in synovial fluid samples in clinical practice

Freise, J.; Meier, Sabine; Zeidler, Henning; Kuipers, Jens G.

doi:10.1007/s00393-015-1589-y

Cited by 8 publications

(4 citation statements)

References 18 publications

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“…The other problem associated with DNA degradation is degradation during sample storage. We tried to solve this issue by replacing the alkali lysis isolation procedure [ 25 , 26 ] by another DNA isolation procedure, using the SiMax™ Genomic DNA Extraction Kit (SBS Genetech, Beijing, China), the QIAamp DSP DNA Genomic Kit (Qiagen, Valencia, CA, USA), and the DNA-sorb B Kit (Amplia s.r.o). However, as in the case of synovial fluid, all silica-based methods dramatically reduced the ability to identify Chlamydia -specific DNA from our sputum samples [ 25 ].…”

Section: Discussionmentioning

confidence: 99%

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Reliable and Sensitive Nested PCR for the Detection of Chlamydia in Sputum

2021

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Chlamydia are Gram-negative, intracellular pathogens colonizing epithelial mucosa. They cause primarily atypical pneumonia and have recently been associated with chronic diseases. Diagnostics relies almost exclusively on serological methods; PCR tests are used rarely because in patients with positive ELISA, it is nearly impossible to identify chlamydial DNA. This paradox is associated with DNA degradation in sputum samples, low abundance, and low sensitivity of PCR systems. In a newly designed and validated “nested” PCR (NPCR) assay, it was possible to amplify DNA of Chlamydia known to infect humans in 31% samples. The reliability of the assay was confirmed by DNA sequencing, and all PCR products belonged exclusively to the Chlamydiales, mainly recognized as Chlamydia pneumoniae. Three samples were related to Ca. Rhabdochlamydia porcellionis and Ca. Renichlamydia lutjani, which infect arthropods. In one case, samples were taken from sick individual, indicating the potential as a human pathogen.

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Section: Discussionmentioning

confidence: 99%

“…The DNA was obtained from sputa according to Freise et al [ 25 , 26 ], with minor modifications. To 0.5 mL of sputa, 200 μL of 50 mM NaOH was added.…”

Section: Methodsmentioning

confidence: 99%

Reliable and Sensitive Nested PCR for the Detection of Chlamydia in Sputum

2021

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“…Interestingly, CT has been detected in human blood samples [46, 47] and shown to infect human peripheral blood cells [48], suggesting that CT may enter and survive in the circulation. This assumption may partially explain the observations that sexually transmitted CT has been detected in synovial tissues [49] and high titers of anti-CT IgG antibodies are detected in women with tubal infertility [50–53]. Regardless how CT gets into women’s GI tract, a burning question is whether the GI tract CT can affect CT pathogenicity in the genital tract [54].…”

Section: Ct Is Frequently Detected In the Gastrointestinal Tractmentioning

confidence: 99%

Chlamydia Spreading from the Genital Tract to the Gastrointestinal Tract – A Two-Hit Hypothesis

Zhong

2018

Trends in Microbiology

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Chlamydia trachomatis, a leading bacterial cause of sexually transmitted infection-induced infertility, is frequently detected in the gastrointestinal tract. Chlamydia muridarum, a model pathogen for investigating C. trachomatis pathogenesis, readily spreads from the mouse genital tract to the gastrointestinal tract, establishing long-lasting colonization. C. muridarum mutants, despite their ability to activate acute oviduct inflammation, are attenuated in inducing tubal fibrosis and are no longer able to colonize the gastrointestinal tract, suggesting that the spread of C. muridarum to the gastrointestinal tract may contribute to its pathogenicity in the upper genital tract. However, gastrointestinal C. muridarum cannot directly autoinoculate the genital tract. Both antigen-specific CD8 T cells and profibrotic cytokines, such as TNFα and IL-13, are essential for C. muridarum to induce tubal fibrosis; this may be induced by the gastrointestinal C. muridarum, as a second hit, to transmucosally convert tubal repairing - initiated by C. muridarum infection of tubal epithelial cells (serving as the first hit) - into pathogenic fibrosis. Testing the two-hit mouse model should both add new knowledge to the growing list of mechanisms by which gastrointestinal microbes contribute to pathologies in extragastrointestinal tissues and provide information for investigating the potential role of gastrointestinal C. trachomatis in human chlamydial pathogenesis.

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“…The development of sensitive and specific multiplex molecular testing methods will be necessary to address the topics raised here, since in contrast to septic arthritis the number of non-culturable bacteria found in the joints of patients with chlamydial and other ReA is usually quite low. In our experience DNA extraction methods and PCR protocols must be adapted to the SF and ST milieu to reach adequate levels of performance [ 51 , 52 , 53 , 54 ]. We assume that research laboratories will develop and test protocols designed for use with joint samples for the identification of Mycoplasma , Ureaplasma , and enteric.…”

Section: Aethiopathogenic and Clinical Implications Of Coinfectionmentioning

confidence: 99%

Coinfection of Chlamydiae and other Bacteria in Reactive Arthritis and Spondyloarthritis: Need for Future Research

Zeidler

Hudson

2016

Microorganisms

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Reactive (inflammatory) arthritis has been known for many years to follow genital infection with the intracellular bacterial pathogen Chlamydia trachomatis in some individuals. Recent studies from several groups have demonstrated that a related bacterium, the respiratory pathogen Chlamydia pneumoniae, can elicit a similar arthritis. Studies of these organisms, and of a set of gastrointestinal pathogens also associated with engendering inflammatory arthritis, have been relatively extensive. However, reports focusing on coinfections with these and/or other organisms, and the effects of such coinfections on the host immune and other systems, have been rare. In this article, we review the extant data regarding infections by multiple pathogens in the joint as they relate to engendering arthritis, and we suggest a number of research areas that must be given a high priority if we are to understand, and therefore to treat in an effective manner, such arthritides.

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Optimized testing for C. trachomatis DNA in synovial fluid samples in clinical practice

Abstract: Alkaline lysis in combination with C. tr.-omp1-152 bp PCR emerged as the most sensitive method for identification of C. tr. in clinical SF samples.

Cited by 8 publications

References 18 publications

Reliable and Sensitive Nested PCR for the Detection of Chlamydia in Sputum

Reliable and Sensitive Nested PCR for the Detection of Chlamydia in Sputum

Chlamydia Spreading from the Genital Tract to the Gastrointestinal Tract – A Two-Hit Hypothesis

Coinfection of Chlamydiae and other Bacteria in Reactive Arthritis and Spondyloarthritis: Need for Future Research

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