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

Optimierte Untersuchung auf C.-trachomatis-DNA in Synovialflüssigkeit für die klinische Praxis

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Abstract

Aim

No standardized polymerase chain reaction (PCR) assay is available for detection of Chlamydia trachomatis (C. tr.) in synovial fluid (SF) for diagnostic use in clinical practice. This study tested the performance of two optimized molecular biology methods, to determine which is best suited for detecting C. tr. in SF clinical samples from patients with various rheumatologic diseases.

Methods

Two DNA extraction methods, i.e., (1) alkaline lysis and (2) QIAEX II Gel Extraction Kit® + cetyltrimethylammonium bromide (CTAB; Qiagen, Hilden, Germany), and C. tr.-omp1-152 bp PCR were tested in SF samples from a total of 329 patients with the following diagnoses: reactive arthritis (ReA; n = 10, 4 patients had posturethritic ReA), undifferentiated arthritis (UA; n = 66), rheumatoid arthritis (RA; n = 169), psoriatic arthritis (PSA; n = 12), and osteoarthritis (OA) n = 72.

Results

In SF samples, C. tr.-omp1-152 bp PCR in combination with alkaline lysis DNA extraction allowed detection of more C. tr.-positive samples: 3/10 (30 %) ReA patients (all with posturethritic ReA) and 20/66 (38 %) UA patients were positive, compared to the 0/10 (0 %) patients with ReA and 1/66 (2 %) with UA detected using the QIAEX II Gel Extraction Kit® + CTAB. Moreover, 2/12 (17 %) SF samples from PSA patients tested positive with alkaline lysis. All samples from patients with OA and RA tested negative.

Conclusion

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.

Zusammenfassung

Ziel

Standardisierte Polymerasekettenreaktions(PCR)-Tests zum Nachweis von Chlamydia trachomatis (C. tr.) in Synovia (SF) für den diagnostischen Einsatz in der klinischen Praxis sind nicht verfügbar. In der vorliegenden Studie wurden 2 optimierte molekularbiologische Verfahren für den Nachweis von C. tr. in SF von Patienten mit unterschiedlichen rheumatischen Erkrankungen geprüft.

Methoden

Zwei DNA-Extraktionsmethoden, 1. alkalische Lyse, 2. QIAex II Gel Extraction Kit® + Cetyltrimethylammoniumbromid (CTAB, Fa. Qiagen, Hilden) mit C.-tr.-omp1-(152 bp-)PCR, wurden zur Analyse von SF-Proben von 329 Patienten mit den folgenden Diagnosen verwendet: reaktive Arthritis (ReA) n = 10 (4 davon mit posturethritischer ReA), undifferenzierte Arthritis (UA) n = 66, rheumatoide Arthritis (RA) n = 169, Psoriasisarthritis (PSA) n = 12, Osteoarthrose (OA) n = 72.

Ergebnisse

Die C.-tr.-omp1-(152 bp-)PCR in Kombination mit alkalischer Lyse erlaubte die häufigere Detektion C.-tr.-positiver SF-Proben: 3/10 (30 %) der ReA-Patienten (alle mit posturethritischer ReA), und 20/66 (38 %) der Patienten mit UA waren positiv – verglichen mit 0/10 (0 %) der ReA- und 1/66 (2 %) der UA-Patienten mit dem QIAmp gel extraction kit® + CTAB. Außerdem waren 2/12 (17 %) der SF-Proben von Patienten mit PSA positiv nach alkalischer Lyse. Alle Proben von Patienten mit OA und RA blieben negativ.

Schlussfolgerung

Alkalische Lyse in Kombination mit C.-tr.-omp1-(152 bp-)PCR zeigte sich als das sensitivste Verfahren zur Detektion von C.-tr. in klinischen SF-Proben.

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Acknowledgements

The authors acknowledge R. Hein, MD, Nienburg, Germany, G. Hoese, MD, Stadthagen, Germany, M. Mahrenholz, MD, Wennigsen, Germany, G. Pott, MD, Hannover, Germany, P. Wagener, MD, Nienburg, Germany and H.-F. Weidemann, MD, Hannover, Germany for contributing synovial fluid samples and M. Rihl, MD, Hannover, Germany for assistance with statistical calculations. We thank Prof Alan P. Hudson, Wayne State University, Detroit, MI, USA for critically reading the manuscript.

This work was supported by grant BMBF rheumatology competence network No. 01 GI 9950; project number C-3.4. The Qiagen products used for evaluation were supplied by courtesy of the Qiagen Company.

Compliance with ethical guidelines

J. Freise, I. Bernau, S. Meier, H. Zeidler, and J.G. Kuipers, state that there are no conflicts of interest.

All studies on humans described in the present manuscript were carried out with the approval of the responsible ethics committee and in accordance with national law and the Helsinki Declaration of 1975 (in its current, revised form). Informed consent was obtained from all patients included in studies.

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Authors and Affiliations

  1. Clinic of Pneumology, Medical School Hannover, Hannover, Germany

    J. Freise

  2. Klinikum Bremen Mitte, Bremen, Germany

    I. Bernau

  3. Clinic of Immunology and Rheumatology, Medical School Hannover, Hannover, Germany

    S. Meier

  4. Emeritus, Medical School Hannover, Hannover, Germany

    H. Zeidler

  5. Div. of Rheumatology, Rotes Kreuz Krankenhaus, St. Pauli Deich 24, 28199, Bremen, Germany

    J.G. Kuipers

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  1. J. Freise
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  2. I. Bernau
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  3. S. Meier
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  4. H. Zeidler
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  5. J.G. Kuipers
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Corresponding author

Correspondence to J.G. Kuipers.

Additional information

Henning Zeidler and Jens G. Kuipers contributed equally to this work.

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Freise, J., Bernau, I., Meier, S. et al. Optimized testing for C. trachomatis DNA in synovial fluid samples in clinical practice. Z Rheumatol 74, 824–828 (2015). https://doi.org/10.1007/s00393-015-1589-y

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  • DOI: https://doi.org/10.1007/s00393-015-1589-y

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